Short sequences (generally about 10 base pairs) of DNA that are complementary to sequences of messenger RNA and allow reverse transcriptases to start copying the adjacent sequences of mRNA. Primers are used extensively in genetic and molecular biology techniques.
Macromolecular molds for the synthesis of complementary macromolecules, as in DNA REPLICATION; GENETIC TRANSCRIPTION of DNA to RNA, and GENETIC TRANSLATION of RNA into POLYPEPTIDES.
The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.
A reverse transcriptase encoded by the POL GENE of HIV. It is a heterodimer of 66 kDa and 51 kDa subunits that are derived from a common precursor protein. The heterodimer also includes an RNAse H activity (RIBONUCLEASE H, HUMAN IMMUNODEFICIENCY VIRUS) that plays an essential role the viral replication process.
An enzyme that synthesizes DNA on an RNA template. It is encoded by the pol gene of retroviruses and by certain retrovirus-like elements. EC
A ribonuclease that specifically cleaves the RNA moiety of RNA:DNA hybrids. It has been isolated from a wide variety of prokaryotic and eukaryotic organisms as well as RETROVIRUSES.
A genus of ciliate protozoa having a dorsoventrally flattened body with widely spaced rows of short bristle-like cilia on the dorsal surface.
Guanine nucleotides which contain deoxyribose as the sugar moiety.
Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.
In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships.
DNA-dependent DNA polymerases found in bacteria, animal and plant cells. During the replication process, these enzymes catalyze the addition of deoxyribonucleotide residues to the end of a DNA strand in the presence of DNA as template-primer. They also possess exonuclease activity and therefore function in DNA repair.
A polynucleotide consisting essentially of chains with a repeating backbone of phosphate and ribose units to which nitrogenous bases are attached. RNA is unique among biological macromolecules in that it can encode genetic information, serve as an abundant structural component of cells, and also possesses catalytic activity. (Rieger et al., Glossary of Genetics: Classical and Molecular, 5th ed)
A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine).
The phosphate esters of DIDEOXYNUCLEOSIDES.
The process by which a DNA molecule is duplicated.
An essential ribonucleoprotein reverse transcriptase that adds telomeric DNA to the ends of eukaryotic CHROMOSOMES.
An antiviral antibiotic produced by Cephalosporium aphidicola and other fungi. It inhibits the growth of eukaryotic cells and certain animal viruses by selectively inhibiting the cellular replication of DNA polymerase II or the viral-induced DNA polymerases. The drug may be useful for controlling excessive cell proliferation in patients with cancer, psoriasis or other dermatitis with little or no adverse effect upon non-multiplying cells.
A single chain of deoxyribonucleotides that occurs in some bacteria and viruses. It usually exists as a covalently closed circle.
Deoxyribonucleic acid that makes up the genetic material of viruses.
A DNA-dependent DNA polymerase characterized in prokaryotes and may be present in higher organisms. It has both 3'-5' and 5'-3' exonuclease activity, but cannot use native double-stranded DNA as template-primer. It is not inhibited by sulfhydryl reagents and is active in both DNA synthesis and repair. EC
The spatial arrangement of the atoms of a nucleic acid or polynucleotide that results in its characteristic 3-dimensional shape.
The monomeric units from which DNA or RNA polymers are constructed. They consist of a purine or pyrimidine base, a pentose sugar, and a phosphate group. (From King & Stansfield, A Dictionary of Genetics, 4th ed)
A characteristic feature of enzyme activity in relation to the kind of substrate on which the enzyme or catalytic molecule reacts.
Inhibitors of reverse transcriptase (RNA-DIRECTED DNA POLYMERASE), an enzyme that synthesizes DNA on an RNA template.
The rate dynamics in chemical or physical systems.
Ribonucleic acid that makes up the genetic material of viruses.
Any detectable and heritable change in the genetic material that causes a change in the GENOTYPE and which is transmitted to daughter cells and to succeeding generations.
The parts of a macromolecule that directly participate in its specific combination with another molecule.
A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc.
The type species of LENTIVIRUS and the etiologic agent of AIDS. It is characterized by its cytopathic effect and affinity for the T4-lymphocyte.
A multistage process that includes cloning, physical mapping, subcloning, determination of the DNA SEQUENCE, and information analysis.
Deoxyribonucleic acid that makes up the genetic material of bacteria.
Technique that utilizes low-stringency polymerase chain reaction (PCR) amplification with single primers of arbitrary sequence to generate strain-specific arrays of anonymous DNA fragments. RAPD technique may be used to determine taxonomic identity, assess kinship relationships, analyze mixed genome samples, and create specific probes.
Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed)
The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells.
A transfer RNA which is specific for carrying lysine to sites on the ribosomes in preparation for protein synthesis.
The relationships of groups of organisms as reflected by their genetic makeup.
The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION.
Constituent of 30S subunit prokaryotic ribosomes containing 1600 nucleotides and 21 proteins. 16S rRNA is involved in initiation of polypeptide synthesis.
The restriction of a characteristic behavior, anatomical structure or physical system, such as immune response; metabolic response, or gene or gene variant to the members of one species. It refers to that property which differentiates one species from another but it is also used for phylogenetic levels higher or lower than the species.
DNA sequences encoding RIBOSOMAL RNA and the segments of DNA separating the individual ribosomal RNA genes, referred to as RIBOSOMAL SPACER DNA.
The biosynthesis of RNA carried out on a template of DNA. The biosynthesis of DNA from an RNA template is called REVERSE TRANSCRIPTION.
A single-stranded DNA-dependent RNA polymerase that functions to initiate, or prime, DNA synthesis by synthesizing oligoribonucleotide primers. EC 2.7.7.-.
Acrylic acids or acrylates which are substituted in the C-2 position with a methyl group.
The sequential correspondence of nucleotides in one nucleic acid molecule with those of another nucleic acid molecule. Sequence homology is an indication of the genetic relatedness of different organisms and gene function.
An adhesion procedure for orthodontic attachments, such as plastic DENTAL CROWNS. This process usually includes the application of an adhesive material (DENTAL CEMENTS) and letting it harden in-place by light or chemical curing.
The functional hereditary units of BACTERIA.
Laboratory techniques that involve the in-vitro synthesis of many copies of DNA or RNA from one original template.
Dental cements composed either of polymethyl methacrylate or dimethacrylate, produced by mixing an acrylic monomer liquid with acrylic polymers and mineral fillers. The cement is insoluble in water and is thus resistant to fluids in the mouth, but is also irritating to the dental pulp. It is used chiefly as a luting agent for fabricated and temporary restorations. (Jablonski's Dictionary of Dentistry, 1992, p159)
The arrangement of two or more amino acid or base sequences from an organism or organisms in such a way as to align areas of the sequences sharing common properties. The degree of relatedness or homology between the sequences is predicted computationally or statistically based on weights assigned to the elements aligned between the sequences. This in turn can serve as a potential indicator of the genetic relatedness between the organisms.
Polymers made up of a few (2-20) nucleotides. In molecular genetics, they refer to a short sequence synthesized to match a region where a mutation is known to occur, and then used as a probe (OLIGONUCLEOTIDE PROBES). (Dorland, 28th ed)
Synthetic or natural oligonucleotides used in hybridization studies in order to identify and study specific nucleic acid fragments, e.g., DNA segments near or within a specific gene locus or gene. The probe hybridizes with a specific mRNA, if present. Conventional techniques used for testing for the hybridization product include dot blot assays, Southern blot assays, and DNA:RNA hybrid-specific antibody tests. Conventional labels for the probe include the radioisotope labels 32P and 125I and the chemical label biotin.
A heat stable DNA-DIRECTED DNA POLYMERASE from the bacteria Thermus aquaticus. It is widely used for the amplification of genes through the process of POLYMERASE CHAIN REACTION. EC 2.7.7.-.
A technique for identifying individuals of a species that is based on the uniqueness of their DNA sequence. Uniqueness is determined by identifying which combination of allelic variations occur in the individual at a statistically relevant number of different loci. In forensic studies, RESTRICTION FRAGMENT LENGTH POLYMORPHISM of multiple, highly polymorphic VNTR LOCI or MICROSATELLITE REPEAT loci are analyzed. The number of loci used for the profile depends on the ALLELE FREQUENCY in the population.
RNA sequences that serve as templates for protein synthesis. Bacterial mRNAs are generally primary transcripts in that they do not require post-transcriptional processing. Eukaryotic mRNA is synthesized in the nucleus and must be exported to the cytoplasm for translation. Most eukaryotic mRNAs have a sequence of polyadenylic acid at the 3' end, referred to as the poly(A) tail. The function of this tail is not known for certain, but it may play a role in the export of mature mRNA from the nucleus as well as in helping stabilize some mRNA molecules by retarding their degradation in the cytoplasm.
Species- or subspecies-specific DNA (including COMPLEMENTARY DNA; conserved genes, whole chromosomes, or whole genomes) used in hybridization studies in order to identify microorganisms, to measure DNA-DNA homologies, to group subspecies, etc. The DNA probe hybridizes with a specific mRNA, if present. Conventional techniques used for testing for the hybridization product include dot blot assays, Southern blot assays, and DNA:RNA hybrid-specific antibody tests. Conventional labels for the DNA probe include the radioisotope labels 32P and 125I and the chemical label biotin. The use of DNA probes provides a specific, sensitive, rapid, and inexpensive replacement for cell culture techniques for diagnosing infections.
Electrophoresis in which agar or agarose gel is used as the diffusion medium.
Deoxyribonucleic acid that makes up the genetic material of plants.
A variety of simple repeat sequences that are distributed throughout the GENOME. They are characterized by a short repeat unit of 2-8 basepairs that is repeated up to 100 times. They are also known as short tandem repeats (STRs).
The genetic constitution of the individual, comprising the ALLELES present at each GENETIC LOCUS.
Ribonucleic acid in bacteria having regulatory and catalytic roles as well as involvement in protein synthesis.
Single-stranded complementary DNA synthesized from an RNA template by the action of RNA-dependent DNA polymerase. cDNA (i.e., complementary DNA, not circular DNA, not C-DNA) is used in a variety of molecular cloning experiments as well as serving as a specific hybridization probe.
Variation occurring within a species in the presence or length of DNA fragment generated by a specific endonuclease at a specific site in the genome. Such variations are generated by mutations that create or abolish recognition sites for these enzymes or change the length of the fragment.
Widely used technique which exploits the ability of complementary sequences in single-stranded DNAs or RNAs to pair with each other to form a double helix. Hybridization can take place between two complimentary DNA sequences, between a single-stranded DNA and a complementary RNA, or between two RNA sequences. The technique is used to detect and isolate specific sequences, measure homology, or define other characteristics of one or both strands. (Kendrew, Encyclopedia of Molecular Biology, 1994, p503)
Deoxyribonucleic acid that makes up the genetic material of fungi.
Genotypic differences observed among individuals in a population.
The internal resistance of a material to moving some parts of it parallel to a fixed plane, in contrast to stretching (TENSILE STRENGTH) or compression (COMPRESSIVE STRENGTH). Ionic crystals are brittle because, when subjected to shear, ions of the same charge are brought next to each other, which causes repulsion.
A variation of the PCR technique in which cDNA is made from RNA via reverse transcription. The resultant cDNA is then amplified using standard PCR protocols.
A method (first developed by E.M. Southern) for detection of DNA that has been electrophoretically separated and immobilized by blotting on nitrocellulose or other type of paper or nylon membrane followed by hybridization with labeled NUCLEIC ACID PROBES.
A phenotypically recognizable genetic trait which can be used to identify a genetic locus, a linkage group, or a recombination event.
Proteins found in any species of bacterium.
Use of restriction endonucleases to analyze and generate a physical map of genomes, genes, or other segments of DNA.
The degree of similarity between sequences of amino acids. This information is useful for the analyzing genetic relatedness of proteins and species.
The regular and simultaneous occurrence in a single interbreeding population of two or more discontinuous genotypes. The concept includes differences in genotypes ranging in size from a single nucleotide site (POLYMORPHISM, SINGLE NUCLEOTIDE) to large nucleotide sequences visible at a chromosomal level.
The intergenic DNA segments that are between the ribosomal RNA genes (internal transcribed spacers) and between the tandemly repeated units of rDNA (external transcribed spacers and nontranscribed spacers).
Constituent of the 40S subunit of eukaryotic ribosomes. 18S rRNA is involved in the initiation of polypeptide synthesis in eukaryotes.
Cements that act through infiltration and polymerization within the dentinal matrix and are used for dental restoration. They can be adhesive resins themselves, adhesion-promoting monomers, or polymerization initiators that act in concert with other agents to form a dentin-bonding system.
Enzymes that catalyze the template-directed incorporation of ribonucleotides into an RNA chain. EC 2.7.7.-.
A large collection of DNA fragments cloned (CLONING, MOLECULAR) from a given organism, tissue, organ, or cell type. It may contain complete genomic sequences (GENOMIC LIBRARY) or complementary DNA sequences, the latter being formed from messenger RNA and lacking intron sequences.
A group of deoxyribonucleotides (up to 12) in which the phosphate residues of each deoxyribonucleotide act as bridges in forming diester linkages between the deoxyribose moieties.

Activation of systemic acquired silencing by localised introduction of DNA. (1/43244)

BACKGROUND: In plants, post-transcriptional gene silencing results in RNA degradation after transcription. Among tobacco transformants carrying a nitrate reductase (Nia) construct under the control of the cauliflower mosaic virus 35S promoter (35S-Nia2), one class of transformants spontaneously triggers Nia post-transcriptional gene silencing (class II) whereas another class does not (class I). Non-silenced plants of both classes become silenced when grafted onto silenced stocks, indicating the existence of a systemic silencing signal. Graft-transmitted silencing is maintained in class II but not in class I plants when removed from silenced stocks, indicating similar requirements for spontaneous triggering and maintenance. RESULTS: Introduction of 35S-Nia2 DNA by the gene transfer method called biolistics led to localised acquired silencing (LAS) in bombarded leaves of wild-type, class I and class II plants, and to systemic acquired silencing (SAS) in class II plants. SAS occurred even if the targeted leaf was removed 2 days after bombardment, indicating that the systemic signal is produced, transmitted and amplified rapidly. SAS was activated by sense, antisense and promoterless Nia2 DNA constructs, indicating that transcription is not required although it does stimulate SAS. CONCLUSIONS: SAS was activated by biolistic introduction of promoterless constructs, indicating that the DNA itself is a potent activator of post-transcriptional gene silencing. The systemic silencing signal invaded the whole plant by cell-to-cell and long-distance propagation, and reamplification of the signal.  (+info)

Impaired translesion synthesis in xeroderma pigmentosum variant extracts. (2/43244)

Xeroderma pigmentosum variant (XPV) cells are characterized by a cellular defect in the ability to synthesize intact daughter DNA strands on damaged templates. Molecular mechanisms that facilitate replication fork progression on damaged DNA in normal cells are not well defined. In this study, we used single-stranded plasmid molecules containing a single N-2-acetylaminofluorene (AAF) adduct to analyze translesion synthesis (TLS) catalyzed by extracts of either normal or XPV primary skin fibroblasts. In one of the substrates, the single AAF adduct was located at the 3' end of a run of three guanines that was previously shown to induce deletion of one G by a slippage mechanism. Primer extension reactions performed by normal cellular extracts from four different individuals produced the same distinct pattern of TLS, with over 80% of the products resulting from the elongation of a slipped intermediate and the remaining 20% resulting from a nonslipped intermediate. In contrast, with cellular extracts from five different XPV patients, the TLS reaction was strongly reduced, yielding only low amounts of TLS via the nonslipped intermediate. With our second substrate, in which the AAF adduct was located at the first G in the run, thus preventing slippage from occurring, we confirmed that normal extracts were able to perform TLS 10-fold more efficiently than XPV extracts. These data demonstrate unequivocally that the defect in XPV cells resides in translesion synthesis independently of the slippage process.  (+info)

Overexpression of CuZn superoxide dismutase protects RAW 264.7 macrophages against nitric oxide cytotoxicity. (3/43244)

Initiation of nitric oxide (NO.)-mediated apoptotic cell death in RAW 264.7 macrophages is associated with up-regulation of mitochondrial manganese superoxide dismutase (MnSOD; SOD2) and down-regulation of cytosolic copper zinc superoxide dismutase (CuZnSOD; SOD1) at their individual mRNA and protein levels. To evaluate the decreased CuZnSOD expression and the initiation of apoptosis we stably transfected macrophages to overexpress human CuZnSOD. Individual clones revealed a 2-fold increase in CuZnSOD activity. Expression of a functional and thus protective CuZnSOD was verified by attenuated superoxide (O2(.)-)-mediated apoptotic as well as necrotic cell death. In this study we showed that SOD-overexpressing macrophages (R-SOD1-12) were also protected against NO.-initiated programmed cell death. Protection was substantial towards NO. derived from exogenously added NO donors or when NO. was generated by inducible NO synthase activation, and was evident at the level of p53 accumulation, caspase activation and DNA fragmentation. Stimulation of parent and SOD-overexpressing cells with a combination of lipopolysaccharide and murine interferon gamma produced equivalent amounts of nitrite/nitrate, which ruled out attenuated inducible NO. synthase activity during protection. Because protection by a O2(.)--scavenging system during NO. -intoxication implies a role of NO. and O2(.)- in the progression of cell damage, we used uric acid to delineate the role of peroxynitrite during NO.-elicited apoptosis. The peroxynitrite scavenger uric acid left S-nitrosoglutathione or spermine-NO-elicited apoptosis unaltered, blocking only 3-morpholinosydnonimine-mediated cell death. As a result we exclude peroxynitrite from contributing, to any major extent, to NO. -mediated apoptosis. Therefore protection observed with CuZnSOD overexpression is unlikely to stem from interference with peroxynitrite formation and/or action. Unequivocally, the down-regulation of CuZnSOD is associated with NO. cytotoxicity, whereas CuZnSOD overexpression protects macrophages from apoptosis.  (+info)

Co-expression of glutathione S-transferase with methionine aminopeptidase: a system of producing enriched N-terminal processed proteins in Escherichia coli. (4/43244)

We describe here an Escherichia coli expression system that produces recombinant proteins enriched in the N-terminal processed form, by using glutathione S-transferase cGSTM1-1 and rGSTT1-1 as models, where c and r refer to chick and rat respectively. Approximately 90% of the cGSTM1-1 or rGSTT1-1 overexpressed in E. coli under the control of a phoA promoter retained the initiator methionine residue that was absent from the mature isoenzymes isolated from tissues. The amount of initiator methionine was decreased to 40% of the expressed cGSTM1-1 when the isoenzyme was co-expressed with an exogenous methionine aminopeptidase gene under the control of a separate phoA promoter. The recombinant proteins expressed were mainly methionine aminopeptidase. The yield of cGSTM1-1 was decreased to 10% of that expressed in the absence of the exogenous methionine aminopeptidase gene. By replacing the phoA with its natural promoter, the expression of methionine aminopeptidase decreased drastically. The yield of the co-expressed cGSTM1-1 was approx. 60% of that in the absence of the exogenous methionine aminopeptidase gene; approx. 65% of the initiator methionine residues were removed from the enzyme. Under similar conditions, N-terminal processing was observed in approx. 70% of the recombinant rGSTT1-1 expressed. By increasing the concentration of phosphate in the growth medium, the amount of initiator methionine on cGSTM1-1 was decreased to 14% of the overexpressed isoenzymes, whereas no further improvement could be observed for rGSTT1-1. The initiator methionine residue does not affect the enzymic activities of either cGSTM1-1 or rGSTT1-1. However, the epoxidase activity and the 4-nitrobenzyl chloride-conjugating activity of the purified recombinant rGSTT1-1 are markedly higher that those reported recently for the same isoenzyme isolated from rat livers.  (+info)

Differential regulation of the human nidogen gene promoter region by a novel cell-type-specific silencer element. (5/43244)

Transfection analyses of the human nidogen promoter region in nidogen-producing fibroblasts from adult skin revealed multiple positive and negative cis-acting elements controlling nidogen gene expression. Characterization of the positive regulatory domains by gel mobility-shift assays and co-transfection studies in Drosophila SL2 cells unequivocally demonstrated that Sp1-like transcription factors are essential for a high expression of the human nidogen gene. Analysis of the negative regulatory domains identified a novel silencer element between nt -1333 and -1322, which is bound by a distinct nuclear factor, by using extracts from adult but not from embryonal fibroblasts. In embryonal fibroblasts, which express significantly higher amounts of nidogen mRNA as compared with adult fibroblasts, this inhibitory nidogen promoter region did not affect nidogen and SV40 promoter activities. The silencer element seems to be active only in nidogen-producing cells. Therefore this regulatory element might function in vivo to limit nidogen gene expression in response to external stimuli. However, none of the identified regulatory elements, including the silencer, contribute significantly to cell-specific expression of the human nidogen gene. Instead we provide evidence that gene expression in epidermal keratinocytes that are not producing nidogen is repressed by methylation-specific and chromatin-dependent mechanisms.  (+info)

Role of retinoid receptors in the regulation of mucin gene expression by retinoic acid in human tracheobronchial epithelial cells. (6/43244)

To investigate which retinoid receptors are critical in the regulation by all-trans-retinoic acid (RA) of the mucin genes MUC2, MUC5AC and MUC5B in cultured normal human tracheobronchial epithelial (NHTBE) cells, we used pan-RAR-, pan-RXR- and RAR- isotype (alpha, beta and gamma)-selective agonists and RARalpha- and RARgamma-selective antagonists (RAR is RA receptor and RXR is retinoid X receptor). RAR-, RARalpha- and RARgamma-selective agonists strongly induced mucin mRNAs in a dose-dependent manner, while the RARbeta-selective retinoid only weakly induced mucin gene expression at very high concentrations (1 microM). The pan-RXR-selective agonist by itself did not induce mucin gene expression, but acted synergistically with suboptimal concentrations of the pan-RAR agonist. A retinoid with selective anti-activator-protein-1 activity only marginally induced mucin gene expression. The RARalpha antagonist strongly inhibited mucin gene induction and mucous cell differentiation caused by RA and by the RARalpha- and RARgamma-selective retinoids. In contrast, the RARgamma antagonist only weakly inhibited RARalpha-selective-retinoid-induced mucin gene expression, but completely blocked mucin gene expression induced by the RARgamma-selective retinoid. Our studies indicate that RARalpha is the major retinoid receptor subtype mediating RA-dependent mucin gene expression and mucous cell differentiation, but that the RARgamma isotype can also induce mucin genes. Furthermore these studies suggest that RARbeta is probably not (directly) involved in RA-induced mucin gene expression.  (+info)

Differential regulation of vascular endothelial growth factor and its receptor fms-like-tyrosine kinase is mediated by nitric oxide in rat renal mesangial cells. (7/43244)

Under conditions associated with local and systemic inflammation, mesangial cells and invading immune cells are likely to be responsible for the release of large amounts of nitric oxide (NO) in the glomerulus. To further define the mechanisms of NO action in the glomerulus, we attempted to identify genes which are regulated by NO in rat glomerular mesangial cells. We identified vascular endothelial growth factor (VEGF) and its receptor fms-like tyrosine kinase (FLT-1) to be under the regulatory control of exogenously applied NO in these cells. Using S-nitroso-glutathione (GSNO) as an NO-donating agent, VEGF expression was strongly induced, whereas expression of its FLT-1 receptor simultaneously decreased. Expressional regulation of VEGF and FLT-1 mRNA was transient and occurred rapidly within 1-3 h after GSNO treatment. Expression of a second VEGF-specific receptor, fetal liver kinase-1 (FLK-1/KDR), could not be detected. The inflammatory cytokine interleukin-1beta mediated a moderate increase in VEGF expression after 24 h and had no influence on FLT-1 expression. In contrast, platelet-derived growth factor-BB and basic fibroblast growth factor had no effect on VEGF expression, but strongly induced FLT-1 mRNA levels. Obviously, there is a differential regulation of VEGF and its receptor FLT-1 by NO, cytokines and growth factors in rat mesangial cells.  (+info)

Comparison of the fibrin-binding activities in the N- and C-termini of fibronectin. (8/43244)

Fibronectin (Fn) binds to fibrin in clots by covalent and non-covalent interactions. The N- and C-termini of Fn each contain one non-covalent fibrin-binding site, which are composed of type 1 (F1) structural repeats. We have previously localized the N-terminal site to the fourth and fifth F1 repeats (4F1.5F1). In the current studies, using proteolytic and recombinant proteins representing both the N- and C-terminal fibrin-binding regions, we localized and characterized the C-terminal fibrin-binding site, compared the relative fibrin-binding activities of both sites and determined the contribution of each site to the fibrin-binding activity of intact Fn. By fibrin-affinity chromatography, a protein composed of the 10F1 repeat through to the C-terminus of Fn (10F1-COOH), expressed in COS-1 cells, and 10F1-12F1, produced in Saccharomyces cerevisiae, displayed fibrin-binding activity. However, since 10F1 and 10F1.11F1 were not active, the presence of 12F1 is required for fibrin binding. A proteolytic fragment of 14.4 kDa, beginning 14 residues N-terminal to 10F1, was isolated from the fibrin-affinity matrix. Radio-iodinated 14.4 kDa fibrin-binding peptide/protein (FBP) demonstrated a dose-dependent and saturable binding to fibrin-coated wells that was both competitively inhibited and reversed by unlabelled 14.4 kDa FBP. Comparison of the fibrin-binding affinities of proteolytic FBPs from the N-terminus (25.9 kDa FBP), the C-terminus (14.4 kDa) and intact Fn by ELISA yielded estimated Kd values of 216, 18 and 2.1 nM, respectively. The higher fibrin-binding affinity of the N-terminus was substantiated by the ability of both a recombinant 4F1.5F1 and a monoclonal antibody (mAb) to this site to maximally inhibit biotinylated Fn binding to fibrin by 80%, and by blocking the 90% inhibitory activity of a polyclonal anti-Fn, by absorption with the 25.9 kDa FBP. We propose that whereas the N-terminal site appears to contribute to most of the binding activity of native Fn to fibrin, the specific binding of the C-terminal site may strengthen this interaction.  (+info)

Chimeric primers: some DNA bases in the primer are replaced with RNA bases, creating a chimeric sequence. The melting ... Integrated DNA Technologies. "Primer design. What is the primer-dimer?". YouTube video. Archived from the original on 2021-12- ... into the primer. The SAMRS DNA could bind to natural DNA, but not to other members of the same SAMRS species. For example, T* ... primers build from SAMRS could avoid primer-primer interactions and allowing sensitive SNP detection as well as multiplex PCR. ...
Disease Primers. 3 (1): 17065. doi:10.1038/nrdp.2017.65. PMID 28960184. S2CID 583204. Abugable AA, Morris JL, Palminha NM, ... Mobile DNA. 9 (1): 15. doi:10.1186/s13100-018-0120-9. PMC 5930866. PMID 29743957. Function of Repetitive DNA DNA+Repetitious+ ... Both types of myotonic dystrophy are due to expanded DNA sequences. In DM1 the DNA sequence that is expanded is CCG while in ... Inverted repeats can play structural roles in DNA and RNA by forming stem loops and cruciforms. For humans, some repeated DNA ...
Nat Rev Dis Primers. 1: 15016. doi:10.1038/nrdp.2015.16. PMC 5381807. PMID 27188665. O'Leary JJ, Kennedy MM, McGee JO (February ... A DNA virus is a virus that has a genome made of deoxyribonucleic acid (DNA) that is replicated by a DNA polymerase. They can ... DNA viruses constitute two Baltimore groups: Group I: double-stranded DNA viruses, and Group II: single-stranded DNA viruses. ... called double-stranded DNA (dsDNA) viruses, and those that have one strand of DNA in their genome, called single-stranded DNA ( ...
The removal of the RNA primer allows DNA ligase to ligate the DNA-DNA nick between the new fragment and the previous strand. ... DNA polymerase III arrives at the RNA primer and begins replicating the DNA, adding onto the 3'OH of the primer ... Because DNA synthesis cannot start de novo, an RNA primer, complementary to part of the single-stranded DNA, is synthesized by ... X can also mediate the switch from RNA primer to DNA. DNA polymerase III synthesizes base pairs at a rate of around 1000 ...
RPR makes use of random primers. These random primers are annealed to template DNA and are then extended by the Klenow fragment ... This is followed by a PCR without primers. In the PCR, DNA fragments with sufficiently overlapping sequences will anneal to ... Since DNA shuffling enables the recombination of genes, protein activities can be enhanced. For example, DNA shuffling has been ... DNA shuffling allows for the fabrication of retroviral vectors with these attributes. For example, DNA shuffling with molecular ...
"Primers, 16S ribosomal DNA - François Lutzoni's Lab". Archived from the original on 2012-12-27. Eden PA, ... primers. Mitochondrial and chloroplastic rRNA are also amplified. The most common primer pair was devised by Weisburg et al. ( ... The two primers are almost identical, but 27F has an M instead of a C. AGAGTTTGATCMTGGCTCAG compared with 8F. In addition to ... James, Greg (15 May 2018). "Universal Bacterial Identification by PCR and DNA Sequencing of 16S rRNA Gene". PCR for Clinical ...
Finally, the strand displacing polymerase begins DNA synthesis where the primer has bound to the target DNA. By using two ... for rapid detection of viral genomic DNA or RNA, pathogenic bacterial genomic DNA, as well as short length aptamer DNA. The ... "Rapid PCR". Retrieved 2014-06-21. "PCR primers work using standard RPA reagents". TwistDx. Retrieved 2015-10-19.[ ... Recombinases are capable of pairing oligonucleotide primers with homologous sequence in duplex DNA. SSB bind to displaced ...
Daisy chaining is when DNA undergoing PCR amplification forms tangles that resemble a 'daisy chain.' During PCR, primers or ... It occurs when DNA undergoing PCR amplification forms tangles that resemble a 'daisy chain.' In essence it teaches DNA to count ... since the denaturing and annealing processes will still continue without primers, the single-stranded DNA molecules will ... Daisy chaining DNA is a form of gene editing, or "gene drive", which, unlike CRISPR, is self limiting. This means that any ...
Unlike S1 mapping, however, primer extension can only be used to locate the 5'-end of an mRNA transcript because the DNA ... The hybridization probe for primer extension is a synthesized oligonucleotide, whereas S1 mapping requires isolation of a DNA ... preferably by using the same primer on the DNA template strand. The exact nucleotide by which the transcription starts at can ... The primer is allowed to anneal to the RNA and reverse transcriptase is used to synthesize cDNA from the RNA until it reaches ...
Universal matK primers can be used for DNA barcoding of angiosperms. LtrA, an open reading frame found in the Lactococcus ... Jing YU, Jian-Hua XU, Shi-Liang ZH (May 2011). "New universal matK primers for DNA barcoding angiosperms". Journal of ...
Crawford, Blair (2015-02-19). "A primer on DNA forensics". Ottawa Citizen. Retrieved 2020-03-03. "Government Bill (House of ... The National DNA Data Bank of Canada (NDDB) is a national DNA Database that was set up in 2000. Managed by the RCMP, it provide ... National DNA Data Bank National Missing Persons DNA Program (Articles with short description, Short description matches ... The database hold 622,478 DNA profiles as of March 31, 2022. The first DNA analysis in Canada for investigative purposes was in ...
Primosomes gives RNA primers for DNA synthesis to strands.[citation needed] ΦX174 is closely related to other microviridae, ... The phi X 174 (or ΦX174) bacteriophage is a single-stranded DNA (ssDNA) virus that infects Escherichia coli, and the first DNA- ... Nobel prize winner Arthur Kornberg used ΦX174 as a model to first prove that DNA synthesized in a test tube by purified enzymes ... H protein (or the DNA Pilot Protein) pilots the viral genome through the bacterial membrane of E.coli bacteria most likely via ...
"Can DNA-Based Ecosystem Assessments Quantify Species Abundance? Testing Primer Bias and Biomass--Sequence Relationships with an ... Consortium for the Barcode of Life Algae DNA barcoding DNA Barcoding DNA barcoding in diet assessment Fish DNA barcoding ... DNA metabarcoding is a method of DNA barcoding that uses universal genetic markers to identify DNA of a mixture of organisms. ... Microbial DNA barcoding is the use of DNA metabarcoding to characterize a mixture of microorganisms. ...
Excision of gene coding sequences from genomic DNA. "BRENDA:". Eun, HM (1996). "Nucleases". Enzymology primer for ... An excess of the enzyme is required to degrade double-stranded DNA or RNA and DNA-RNA hybrids, and in this case, AT-rich ... Nuclease MB is a specific DNA and RNA exo-endonuclease which will degrade single-stranded extensions from the ends of DNA and ... The enzyme degrades single-stranded DNA or RNA to nucleoside 5'-monophosphates, but does not digest double-stranded DNA, double ...
... was the first publicly available software for DNA primer design. The first papers describing ... Oligo Primer Analysis Software by Molecular Biology Insights FastPCR Gene Designer Geneious MacVector Netprimer Primer Premier ... "Oligo Primer Analysis Software from Molecular Biology Insights". Wojciech Rychlik (1993) Selection of Primers for Polymerase ... Wojciech Rychlik (2007). "OLIGO 7 Primer Analysis Software". PCR Primer Design. Methods in Molecular Biology. Vol. 402. pp. 35- ...
DNA polymorphisms amplified by arbitrary primers are useful genetic markers. Nucleic Acids Research. 18, 6531-6535 (1990). ... as well as on DNA-DNA hybridization assays and phenotypic characterizations. The type strain EN-119T was isolated from ... On the basis of DNA relatedness, both organisms could be included in a single taxon. However, the CDC enteric group 69 was ... 8, 281-289 (2002). Ruiz J. Mechanisms of resistance to quinolones: target alterations, decreased accumulation and DNA gyrase ...
Two sets of primers are used in two successive reactions. In the first PCR, one pair of primers is used to generate DNA ... January 1988). "Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase". Science. 239 (4839): 487-91 ... Two amplifications are then carried out on the bisulfite-treated DNA: One primer set anneals to DNA with cytosines ( ... uses a recombinase to specifically pair primers with double-stranded DNA on the basis of homology, thus directing DNA synthesis ...
Caramanica, Jon (February 7, 2018). "A K-Pop Primer for Olympic Listening". The New York Times. Archived from the original on ... 뮤직뱅크' 방탄소년단 'DNA' 연이은 1위 행진 4관왕…케이윌-B1A4 컴백무대 ['Music Bank' BTS 'DNA' 1st march four crowns… K.Will-B1A4 Comeback Stage]. Busan ... DNA - 방탄소년단 [DNA - BTS]. Melon. Archived from the original on September 22, 2017. Retrieved August 3, 2020. Boyle, Kelly ( ... Ahn, Tae-Hyun (September 24, 2017). '인기가요' 방탄소년단, 역대급 컴백무대로 증명한 우월 'DNA' ['Inkigayo' BTS, superior 'DNA' proved to be the best ...
Evans, Nathan T.; Lamberti, Gary A. (January 2018). "Freshwater fisheries assessment using environmental DNA: A primer on the ... DNA barcoding DNA barcoding in diet assessment Algae DNA barcoding Microbial DNA barcoding Aquatic macroinvertebrate DNA ... DNA barcoding methods for fish are used to identify groups of fish based on DNA sequences within selected regions of a genome. ... Primer design is crucial for metabarcoding success. Some studies on primer development have described cytochrome B and 16S as ...
Universal primer cocktails for fish DNA barcoding. Molecular Ecology Notes. 7, 544-54. Prince C. 2009. Practical Manual of ...
A genetic marker is a gene or DNA sequence with a known location on a chromosome that can be used to identify individuals or ... the development and use of evolutionarily conserved sets of PCR primers. the use of microsatellite loci that vary among ... The marker could be a short DNA sequence, such as a sequence surrounding a single base-pair change, known as a single ... the development of advanced DNA sequencing techniques. Many things utilized for studying larger organisms has not been possible ...
The amount of DNA produced in LAMP is considerably higher than PCR-based amplification. Primer design could be performed using ... The larger number of primers per target in LAMP increases the likelihood of primer-primer interactions for multiplexed target ... Because LAMP uses 4 (or 6) primers targeting 6 (or 8) regions within a fairly small segment of the genome, and because primer ... open-source or commercial software packages are generally used to assist with LAMP primer design, although the primer design ...
"DNA Polymorphisms Amplified by Arbitrary Primers Are Useful as Genetic Markers". Nucleic Acids Research. 18 (22): 6531-6535. ... Books Kidwell, K. K.; Osborn, T. C. (1992). "Simple Plant DNA Isolation Procedures". In Beckman, J. S.; Osborn, T. C. (eds.). ... Doyle, J. J.; Doyle, J. L. (1987). "A Rapid DNA Isolation Procedure for Small Quantities of Fresh Leaf Tissue". Phytochemical ... Sharma, R.; Mahla, R. H.; Mohapatra, T.; Bhargava, C. S.; Shama, M. M. (2003). "Isolating Plant Genomic DNA Without Liguid ...
"Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase". Science. 239 (4839): 487-491. Bibcode: ... DNA, based on A-T, C-G base pairs, are formed in well-aligned sequences. Through precise sequences of DNA, 20 amino acids are ... At first, taking DNA or RNA as templates, scientists developed a series of peptide nucleic acid (PNA)-based polymers, without ... In nature, DNA, RNA, proteins and other macromolecules can also be recognized as sequence-controlled polymers for their well- ...
of the domestic fowl using DNA polymorphisms amplified by arbitrary primers". Parasitology Research. 79 (2): 98-102. doi: ... DNA assays and recombinant DNA techniques. PCR has proven most useful for outbreak surveillance. Prior to these methods, ...
MethPrimerDB Contains 259 primer sets from human, mouse and rat for DNA methylation analysis. The Histone Database Contains 254 ... "methPrimerDB: the DNA methylation analysis PCR primer database". Archived from the original on 2014-07-15. Retrieved 2010-01-29 ... MethyLogiX DNA methylation database Contains DNA methylation data of human chromosomes 21 and 22, male germ cells and late- ... The Krembil Family Epigenetics Laboratory Contains DNA methylation data of human chromosomes 21, 22, male germ cells and DNA ...
2017). Forensic DNA analysis : a primer for courts. London: Royal Society. ISBN 978-1-78252-301-7. OCLC 1039675621. Bowcott, ... A national DNA database is a DNA database maintained by the government for storing DNA profiles of its population. Each DNA ... A DNA database or DNA databank is a database of DNA profiles which can be used in the analysis of genetic diseases, genetic ... As the DNA profiles can be stored indefinitely in DNA database, it has raised concerns that these DNA samples can be used for ...
DNA polymerase is then able to add DNA nucleotides to the RNA primer and thus begin the process of constructing a new ... The DNA single-strand template serves to guide the synthesis of a complementary strand of DNA. DNA replication begins at a ... Nuclear DNA and mitochondrial DNA differ in many ways, starting with location and structure. Nuclear DNA is located within the ... Nuclear DNA is diploid, ordinarily inheriting the DNA from two parents, while mitochondrial DNA is haploid, coming only from ...
2017). Forensic DNA analysis : a primer for courts. London: Royal Society. ISBN 978-1-78252-301-7. OCLC 1039675621. "DNA ... National Police Improvement Agency, NPIA and the DNA Database The national DNA database, Home Office "The DNA Expansion ... "Has our DNA database gone too far?", BBC News Give Us Your DNA, BBC Panorama Guardian Podcast on the pros and cons of the DNA ... DNA profiling system (SGM+ DNA profiling system since 1998). All data held on the National DNA Database is governed by a tri- ...
These tools are used to optimize the design of primers for target DNA or cDNA sequences. Primer optimization has two goals: ... The last 10-12 bases at the 3' end of a primer are sensitive to initiation of polymerase extension and general primer stability ... On the other hand, FastPCR, a commercial application, allows simultaneous testing of a single primer or a set of primers ... nor should the primer pairs bind to conserved regions of a gene family. If the selectivity is poor, a set of primers will ...
Funk's research included detailing evolutionary relationships and biogeography using plant DNA. She co-discovered the ... "The compleat cladist: A primer of phylogeny procedures". ...
1976: Harald zur Hausen and Gisam found HPV DNA in cervical cancer and genital warts; Hausen later won the Nobel Prize for his ... Gottlieb N (24 April 2002). "A Primer on HPV". Benchmarks. National Cancer Institute. Archived from the original on 26 October ... Dürst M, Gissmann L, Ikenberg H, zur Hausen H (June 1983). "A papillomavirus DNA from a cervical carcinoma and its prevalence ... A description of human papillomavirus (HPV) by electron microscopy was given in 1949, and HPV-DNA was identified in 1963. It ...
While Harjo refers to "Native DNA", there is no DNA test that can reliably confirm Native American ancestry, and no DNA test ... A Quick Primer for Reporters and Others", by Kim TallBear (Sisseton-Wahpeton) "Playing Pretendian", Code Switch, NPR ... The commercial DNA companies that offer ethnicity tests do not have a large enough pool of North American DNA to provide ... DNA tests are setting up other problems involving those who discover Native DNA [sic] in their bloodline. When individuals ...
... consequently requiring many primers. The RNA primers of Okazaki fragments are subsequently degraded by RNase H and DNA ... of either parent DNA or newly formed DNA and thereafter the ligating activity ligates that broken DNA strand and so the two DNA ... which will continue to unwind the DNA as the DnaG primase lays down an RNA primer and DNA Polymerase III holoenzyme begins ... DNA polymerase III holoenzyme is loaded into the DNA and replication begins. The catalytic mechanism of DNA polymerase III ...
"Panamá confirma el primer caso de viruela del mono". TVN Noticias (in Spanish). 5 July 2022. Retrieved 5 July 2022. "2022 U.S. ... Diagnosis can be confirmed by testing a lesion for the virus's DNA. There is no known cure. A study in 1988 found that the ... "Detectan el primer caso de viruela del mono en México". Proceso (in Spanish). Archived from the original on 28 May 2022. ... "Honduras registra primer caso de viruela del mono en hombre menor de 50 años". Swissinfo (in Spanish). Retrieved 13 August 2022 ...
... or DNA. There are meant to be certain cells in a specific area, for the pineal region these are ependymal cells, and the cells ... A Primer of Brain Tumors (2nd ed.). Des Plaines: American Brain Tumor Association. (Orphaned articles from December 2021, All ...
This is the first dataset related to Monkeypox viral DNA in wastewater in Bangkok. Monkeypox viral DNA was first detected in ... "MINSA confirma primer caso de la viruela del mono en el Perú" [MINSA confirms first case of monkeypox in Peru]. Ministry of ... "MINSAL confirma el primer caso de Viruela del Mono en Chile" [MINSAL confirms the first case of Monkeypox in Chile]. Ministry ... "Confirman primer caso de viruela del mono en Bolivia" [First case of monkeypox confirmed in Bolivia]. El Deber (in Spanish). 1 ...
Yan, Hong; Kun, Liu Xiao (13 Jun 2013). "5. Real-Time Fluorescent PCR by Labeled Primer with a Single Fluorescent Molecule". In ... DNA analysis and PCR. Its products leverage the capabilities of multilayer soft lithography to create microfluidic devices: ... The company's second marketed product targeted high-throughput DNA amplification and was launched in 2006 under the brand " ...
If the DNA is damaged, p53 will either repair the DNA or trigger the apoptosis of the cell. If p53 is dysfunctional or mutated ... This article incorporates public domain material from Science Primer. NCBI. Archived from the original on 8 December 2009. ... DNA replication occurs during the C period. The D period refers to the stage between the end of DNA replication and the ... It is estimated that in normal human cells about 1% of single-strand DNA damages are converted to about 50 endogenous DNA ...
The DNA analysis of contemporary persons from this area shows maternal ancestry from the Mandinka, Wolof, and Fulani peoples ... que se usaba ya en el primer tercio de siglo xvi, y que ha venido a resultar otro de los numerosos antillanismos que la ... "African DNA Project mtDNA Haplogroup L1b". 8 May 2008. Archived from the original on 8 May 2008.{{cite web}}: CS1 maint: bot: ...
A pilot study of mitochondrial DNA and Y-DNA". Canadian Journal of Gastroenterology. 25 (6): 324-326. doi:10.1155/2011/463810. ... Disease Primers. 4: 18016. doi:10.1038/nrdp.2018.16. PMC 7775623. PMID 29620054. > hemochromatosis, ... DNA/screening: the current standard of practice in diagnosis of hemochromatosis, places emphasis on genetic testing. Positive ...
Pérez, José I. (19 August 2017). "El ADN del Atlético no se ficha" [You can't sign Atlético's DNA]. Marca (in Spanish). ... Roma, Raimon (22 July 2017). "Christian Stuani és el primer davanter" [Christian Stuani is the first forward]. Diari de Girona ...
... and for his editing of the Primers page for Texter received his secondary education at Penn Manor High ... initiating Monte Carlo analyses and modeling of DNA photochemical processes, and a postdoctoral year with Eugene S. Stevens at ... "Saturation photodimerization of thymines in DNA". Biopolymers. 30 (7-8): 797-802. doi:10.1002/bip.360300714. PMID 2275979. ... ". "Find a Conference". "National Merit® Scholarship Program". Miner, Dolores (1991). "Former ...
Except for one study in Europe, much of the data implicating lizards is based on DNA detection of the spirochete and has not ... Disease Primers. 2: 16090. doi:10.1038/nrdp.2016.90. PMC 5539539. PMID 27976670. "Lyme borreliosis" (PDF). ECDC. Archived (PDF ... The 2010 autopsy of Ötzi the Iceman, a 5,300-year-old mummy, revealed the presence of the DNA sequence of Borrelia burgdorferi ... Polymerase chain reaction (PCR) tests for Lyme disease have also been developed to detect the genetic material (DNA) of the ...
Argüelles, José; Argüelles, Lloydine (1992). Thirteen Moons in Motion: A Dreamspell Primer. Portland, OR: Planet Art Network. ... the 64-unit DNA code, and many other "divinatory" systems, including the cosmology of Ibn al-Arabi of the 28 lunar mansions and ...
Certain polymerase chain reaction (PCR) primers have been designed and tested to consistently identify S. perseae with DNA ...
The transferred DNA is piloted to the plant cell nucleus and integrated into the host plants genomic DNA.The plasmid T-DNA is ... Gibson G, Muse SV (2009). A Primer of Genome Science (Third ed.). 23 Plumtree Rd, Sunderland, MA 01375: Sinauer Associates. pp ... The solution, along with the DNA, is encapsulated by the cells and a small amount of DNA can be integrated into the genome. ... In plants the DNA is often inserted using Agrobacterium-mediated recombination, taking advantage of the Agrobacteriums T-DNA ...
And a primer sequence at the end, it is a sequence whose design varies and is what will allow the design of primers and ... Pairs of probes are hybridized to the sample DNA, with each probe pair designed to query for the presence of a particular DNA ... In a standard multiplex PCR reaction, each fragment needs a unique amplifying primer pair. These primers being present in a ... The process consists of multiple steps: The sample DNA is denatured, resulting in single-stranded sample DNA. ...
... also has advanced tools for finishing tasks like automated primer picking Phred Phrap Gordon D, Abajian C, Green P (1998 ... Consed is a program for viewing, editing, and finishing DNA sequence assemblies. Originally developed for sequence assemblies ... and finishing tasks like primer picking and gap closure. Development of Consed has continued after the completion of the Human ...
Repetitive DNA sequences must be blocked by adding short fragments of DNA to the sample. The probe is then applied to the ... Bacterial FISH probes are often primers for the 16s rRNA region. FISH is widely used in the field of microbial ecology, to ... Preparing DNA probes for one species and performing FISH with this probe allows one to visualize the distribution of this ... In biology, a probe is a single strand of DNA or RNA that is complementary to a nucleotide sequence of interest. RNA probes can ...
Two bands of DNA, at 555 bps and 371 bps, are resolved if both the AMELX and AMELY versions of the gene are present (i.e. the ... Mutation in regions of AMELY intron 1 commonly used as primer annealing sites may disable PCR amplification. A 6bp insertion to ... sample is from a male) or one band of DNA, at 555 bps, if the AMELX version only is present (i.e. the sample is from a female ...
This protein is capable of generating the RNA primers required by DNA polymerase gamma to initiate replication of mitochondrial ... DNA and Cell Biology. 16 (9): 1111-22. doi:10.1089/dna.1997.16.1111. PMID 9324313. Li LY, Luo X, Wang X (Jul 2001). " ... DNA and Cell Biology. 34 (2): 92-100. doi:10.1089/dna.2014.2530. PMC 4308826. PMID 25401220. Diener T, Neuhaus M, Koziel R, ... The enzyme encoded by this gene is a member of the conserved DNA/RNA non-specific ββα-Me-finger nuclease family and possesses a ...
Primer design and optimization for RAPD analysis of Nepenthes. Biologia Plantarum 43(1): 153-155. doi:10.1023/A:1026535920714 ... dan amplified ribosomul DNA restriction analysis (ARDRA). MSc thesis, Bogor Agricultural University, Bogor. Yogiara, A. Suwanto ...
Many of such recent studies rely on DNA metabarcoding-high‐throughput sequencing of PCR amplicons using generic primers. Canada ... Relic DNA dynamics Extracellular DNA, sometimes called relic DNA, is DNA from dead microbes. Naked extracellular DNA (eDNA), ... The DNA in the sample is extracted and purified. The purified DNA is then amplified for a specific gene target so it can be ... Extracellular DNA in surface deep-sea sediments is by far the largest reservoir of DNA of the world oceans. The main sources of ...
Wooddell, C I; Burgess, R R (1996). "Use of Asymmetric PCR to Generate Long Primers and Single-stranded DNA for Incorporating ... Asymmetric PCR can be used to form single stranded DNA from double stranded DNA, which is then used for DNA sequencing in the ... limiting primer with a higher melting temperature than the excess primer to maintain reaction efficiency as the limiting primer ... Asymmetric PCR is a variation of PCR used to preferentially amplify one strand of the original DNA more than the other. The ...
DNA - DNA fragmentation - DNA replication - DNA sequence - DNA topology - DNA transposable element - DNA virus - DNA-binding ... primer - prion - progesterone receptor - prokaryote - prolactin - prolactin receptor - proline - promoter - prostaglandin e ... RNA-directed DNA polymerase - rod outer segment - rough ER sarcoplasmic reticulum - satellite DNA - scientific notation - SDS- ... cyclic AMP-responsive DNA-binding protein - cyclic electron flow - cyclic nucleotide - cyclic peptide - cyclin - cyclin A - ...
rRNA gene primers were used to test Neobodo's global distribution and genetic diversity. The non-overlap between environmental ... DOI: 10.1016/j.jembe.2013.03.017 Von Der Heyden, S., and Cavalier-Smith, T. 2005: Culturing and Environmental DNA Sequencing ... They also house discoid shaped mitochondrial cristae and a compact kinetoplast (a DNA-containing granule located within a ... DNA sequences and those from cultures suggests that there are hundreds, possibly thousands, of different rRNA gene sequences of ...
... a credit dispute between universal primer technology and DNA barcoding came to light. Verma has argued that DNA barcoding, a ... Verma was primarily known for his contributions to the development of "universal primer technology", a first generation DNA ... short stretch of DNA from mitochondrial genome, amplified using the specific universal primers, to assign the identity of an ... "ROLE OF DNA FORENSICS IN CURBING ILLEGAL WILDLIFE TRADE" (PDF). WWF-India. p. 14. Archived (PDF) from the original on 30 ...
Of these, only DNA vaccines have entered into clinical trials. Ribavirin may be a drug for HPS and HFRS, but its effectiveness ... for the production of capped primers used to initiate transcription of viral mRNAs. As a result of this cap snatching, the ... Apart from these vaccines, four types of vaccines have been researched: DNA vaccines targeting the M genome segment and the S ...
Certain configurations have been shown to stabilize Watson-Crick antiparallel duplex DNA. TINA-DNA primers have been shown to ... stabilizes Hoogsteen triplex DNA formation from double-stranded DNA (dsDNA) and TFOs. Its ability to twist around a triple bond ... Diagnostic assays using DNA hybridization are limited by the dissociation of antiparallell duplex helices. This can be improved ... Studies show that the greatest increase in stability occurred when intercalating primers were used at the 3' and 5' ends. ...
Trichophyton species DNA is amplified very poorly by the ITS primer set used for most other molds. There is a special set of ... Target Genes, Primer Sets, and Thermocycler Settings for Fungal DNA Amplification. *Anamorph and Teleomorph Names for Candida ... Target Genes, Primer Sets, and Thermocycler Settings for Fungal DNA Amplification. ... This document describes some of the target genes and primers that can be used for DNA sequence-based identification of fungi ...
DNA polymerases that can replicate DNA without a primer! Find all the Institut Pasteur news and projects on its website. ... All previously known DNA polymerases needed a primer, which could come either from another protein or from a pre-existing DNA ... Biotechnology: DNA polymerases that can replicate DNA without a primer!. *Blood test: a potential new tool for controlling ... Biotechnology: DNA polymerases that can replicate DNA without a primer!. *Blood test: a potential new tool for controlling ...
Antibodies / Assay Kits / Biology Cells / cDNA / Clia Kits / Culture Cells / Devices / DNA / DNA Templates / DNA Testing / ... Copyright © 2023 Primers, 16s Ribosomal DNA - François Lutzonis Lab - OnePress theme by FameThemes ...
For DNA sample concentration less than 50 ng/μL; the DNA was concentrated to 70 ng/μL using Glycogen -Sodium acetate (3 M, ... PCR Primers Used in This Study. The PCR primers used in this study are summarized in Table S1. ... DNA Extraction from Low-Melt Agarose Gel and SANGER Sequencing. After DNA fragments were run and separated by agarose gel ... Random primers and gene-specific designed primers were used to enhance the targeting and the amplification of the interested ...
The copies of DNA produced by PCR provide researchers with sufficient copies for other applications in research including ... is a basic method used in molecular biology to produce copies of a small target region of DNA in a sample. The basics to PCR ... For bacterial DNA 10E5 copies will require only 300 picograms of DNA. For human DNA 10E5 will require over 300 nanograms of DNA ... Concentration of the Primers. Primers are the determining factor of what region of the DNA will be amplified by PCR. The ...
Finally, the obtained supernatants were used as DNA source directly.. Primers. Three pairs of primers were designed according ... The primers were synthesised by TIB MOLBIOL (Berlin, Germany). The primers that were used for specific amplification of B. ... primers were used to confirm the presence of plasmids pX01 and pX02 respectively. These primers were confirmed to be specific ... Total DNA extraction. Bacterial isolates were cultured on blood agar plates and then 1 colony was picked and resuspended in ...
We recommend DNA barcoding to be applied to other biodiversity hotspots for quickly and cost-efficiently flagging species ... We successfully DNA-barcoded 380 individuals of all 31 presently recognized species. The specimen identification success rate ... we performed the most comprehensive DNA barcoding study on an island group to date to test its applicability to specimen ... Few DNA barcoding studies of squamate reptiles have been conducted. Due to the significance of the Socotra Archipelago (a ...
DNA Primers / genetics * Down-Regulation * Female * Gene Expression Regulation, Viral * Gene Products, pol / genetics ...
Pick Primers Design and test primers for this sequence using Primer-BLAST. ... Arachis hypogaea DNA, clone: KIAC05C12, genomic survey sequence Arachis hypogaea DNA, clone: KIAC05C12, genomic survey sequence ...
Ampli Taq DNA Polymerase; Perkin Elmer), 10 μl of 10× PCR buffer (Perkin Elmer), and 200 ng of each primer. For reamplification ... of polyclonal tonsillar DNA (B), of DNA from a patient with coeliac disease showing polyclonal amplification (C), of DNA from a ... PRIMER SETS. Oligonucleotide consensus primers for detection of Vγ1 to Vγ8 segments were used in a semi nested polymerase chain ... DNA ISOLATION. DNA was extracted from formalin fixed, paraffin embedded, duodenal and colonic biopsies and resected tumour ...
Proposition 69: DNA Samples. *Required DNA samples to be collected from all convicted felons and certain arrestees for ... We conclude this primer by describing the most significant changes in California criminal justice law over the past two decades ... This primer is organized into different sections that seek to answer key questions about the criminal justice system in ... We note that the crime rates cited in this primer are based on the federal Uniform Crime Reporting program, which is designed ...
Primers are listed in Supplementary Table 1. The Q56T mutant contained an inadvertent mutation, A52V, which mimicked the Val ... Iwahara, J., Wojciak, J. M. & Clubb, R. T. Improved NMR spectra of a protein-DNA complex through rational mutagenesis and the ... Hihara, Y., Kamei, A., Kanehisa, M., Kaplan, A. & Ikeuchi, M. DNA microarray analysis of cyanobacterial gene expression during ... All resulting constructs were verified by DNA sequencing (Evrogen, Moscow, Russia).. Protein production and sample preparation ...
However, the proper primers for Pol  were designed, and the concentrations of MgSO4 and the primers were both optimized at 2mM ... This study continues the analysis of the DNA Pol  subunit by beginning to examine the effect DPH has on the expression of Pol ... Studying the effects of dilantin on DNA polymerase delta RNA expression in second cell cycle preimplantation mouse embryos : ... Studying the effects of dilantin on DNA polymerase delta RNA expression in second cell cycle preimplantation mouse embryos : ...
It is absorbed into our DNA. We could not become professionally qualified until we could convince our peers that we could be ... A Design Primer for Engineers. For a word that can so vastly change the fortunes of a company, its worth noting that no ... This primer is the first step at building a solid bridge between our professions. So, chill. This is not a definitive design ... Furthermore, this primer is being written by an engineer for engineers. While Ive spent a good many years soaking in design, ...
200 nmol/L each primer, 1 × PCR buffer, 4 mmol/L MgCl2, 200 mmol/L dNTPs and 1 U of Hot Star Taq DNA polymerase (Takara, China ... O6-Methylguanine-DNA methyltransferase (MGMT) is a ubiquitous DNA repair protein that can correct the mismatch of O6 alkyl ... Highlights DNA damage was caused by fluorosilicic acid in human osteoblast cells. DNA damage was caused by cotinine in human ... p53-Mediated down-regulation of the human DNA repair gene O6-methylguanine-DNA methyltransferase (MGMT) via interaction with ...
DNA template. X μl. Forward Primer (5 μM)*. 0.75 μl. Reverse Primer (5 μM)*. 0.75 μl. ... Our high quality reagents are available for every workflow, including popular DNA assembly methods such as NEBuilder® HiFi DNA ... Whether you are performing your first cloning experiment or constructing multi-fragment DNA assemblies, NEB® has the solution ...
Primers were designed for these loci. DNA is being extracted from a world-wide collection of 83 isolates. The microsatellite ...
Design and analyze DNA and RNA oligos for insight into behavior and properties. ... OligoAnalyzer is a primer analysis tool for oligonucleotides. ... Enter your primer or other oligo sequence. *Adjust calculation ... 2023 Integrated DNA Technologies, Inc.. Trademarks contained herein are the property of Integrated DNA Technologies, Inc. or ... Your product is now available from Integrated DNA Technologies.. Many of the Swift products you have grown to love are now part ...
Order Ultramer DNA Oligos, long, high-quality oligos for demanding applications such as cloning, ddRNAi, homology-directed ... 1. Skerra A. Phosphorothioate primers improve the amplification of DNA sequences by DNA polymerases with proofreading activity ... 200 pmol Ultramer DNA Oligo. 4 nmol Ultramer DNA Oligo. 20 nmol Ultramer DNA Oligo. PAGE Ultramer DNA Oligo. ... DNA? RNA is inherently less stable than DNA due to its chemical structure. Additionally, RNases are more prevalent in standard ...
Increased specificity of primer annealing.. CoralLoad PCR Buffer.. HotStarTaq Plus procedure.. Principle. HotStarTaq Plus DNA ... Increased specificity of primer annealing.. CoralLoad PCR Buffer.. Applications. HotStarTaq Plus DNA Polymerase is highly ... HotStarTaq Plus DNA Polymerase. HotStarTaq Plus DNA Polymerase. For fast and highly specific amplification in all applications ... HotStarTaqPlus DNA Polymerase. HotStarTaq DNA Polymerase. Hot-start enzyme from Supplier AII. Supplier R. Supplier I (antibody- ...
We then adapted a method and pipeline for COI metabarcoding using generalist primers that target the eukaryote diversity ... DNA metabarcoding techniques allow for fast and comprehensive assessment of biodiversity in both terrestrial and marine ... DNA metabarcoding techniques allow for fast and comprehensive assessment of biodiversity in both terrestrial and marine ... For DNA extraction, 5 g of each homogenate was processed with the PowerMax Soil DNA Isolation Kit (QIAGEN). DNA concentration ...
PCR primers were designed targeting both 5-end and 3-end of the integration sites, using one primer specific to the insert ... Toxicity of plasmid DNA in primary cells is caused by activation of cytosolic DNA sensors that induce subsequent apoptosis and ... This linearized DNA template is then integrated into the host genome, possibly through DNA repair mechanisms related to Single ... et al. Multiple cytosolic DNA sensors bind plasmid DNA after transfection. Nucleic Acids Res. 47, 10235-10246 (2019).. ...
K. Müller 2005b: SeqState: Primer design and sequence statistics for phylogenetic DNA datasets. - Appl. Bio-informatics 4: 65- ... D. Edwards , A. Horn , D. Taylor , V. Savolainen & J. A. Hawkins 2008: DNA barcoding of a large genus, Aspalathus L. (Fabaceae) ... W. J. Kress , K. J. Wurdack , E. A. Zimmer , L. A. Weigt & D. H. Janzen 2005: Use of DNA barcodes to identify flowering plants ... D. Quandt , K. Müller & S. Huttunen 2003: Characterisation of the chloroplast DNA psbT-H region and the influence of dyad ...
Control schemes for microfludic viral DNA/RNA amplification. Talk, Proceedings of the 27th Annual Meeting of the American ... Sensitive, microliter PCR with consensus degenerate primers for Epstein Barr virus amplification. Biomedical Microdevices. 2013 ... Simultaneous amplification of multiple DNA targets with optimized annealing temperatures. Talk, Proceedings of the Biomedical ... Sensitive, microliter PCR with degenerate primers for respiratory virus detection and discovery. Poster presentation, ...
... was performed using a volume of 50 μL containing 30 ng of DNA, 50 pmol of each primer, 2 mM dNTPs and 1.0 U GoTaq DNA ... DNA was isolated from blood samples or from lymphocyte cultures standard methods. Amplification of coding exons, including the ... Samples were separated and analysed on an Applied Biosystems 3730xl DNA Analyser. Sequence data were evaluated using CodonCode ... polymerase (Promega Corporation, Madison, WI, USA). PCR conditions and primer details are available upon request. PCR products ...
Sometimes, certain areas of the exome are incompletely sequenced, for example when DNA primers bind poorly. Johnson and ... The strong bonds between those nucleotides can interfere with sequencing and primer binding due to their high melting ... Paquis-Flucklinger and colleagues reported that their myopathy patients had disorganized mitochondria with fragmented DNA. The ...
Primer design & Analysis tools Multiple primer analyzer TM calculator qPCR qpcr efficiency calculator FAQ(よくある質問と答) ベクターマップ & シ ... Schematic and example of a DNA mismatch detection assay. Genomic DNA (blue) from edited cells contains wild type and edited DNA ... Explore カスタムオリゴ合成 一本鎖RNA/RNA-DNAキメラ合成 カスタムsiRNA合成 カスタムアンチセンスオリゴ合成 Custom DNA カスタムmicroRNA合成 遺伝子解析用標準サンプル 遺伝子解析用標準サンプル show/hide ... Proper assessment of gene editing with DNA mismatch
  • DNA sequences are available from GenBank (accessions KU567304 to KU567683). (
  • Ultramer DNA Oligos are long, single- and double-stranded synthetic DNA sequences. (
  • An RLU measurement equal to or greater than the Cutoff Value indicates the presence of HPV DNA sequences in the specimen. (
  • An RLU measurement less than the Cutoff Value indicates the absence of the specific HPV DNA sequences tested or HPV DNA levels below the detection limit of the assay. (
  • Short sequences (generally about 10 base pairs) of DNA that are complementary to sequences of messenger RNA and allow reverse transcriptases to start copying the adjacent sequences of mRNA. (
  • Prevalences of herpesviruses DNA sequences in salivary gland biopsies from primary and secondary Sjogren's syndrome using degenerated consensus PCR primers. (
  • PCR with oligonucleotide primers to the fistula were diagnosed with tubercular fis- end of IS 6110 . (
  • The discovery of a DNA polymerase that can start DNA synthesis on its own is important from an evolutionary and biotechnological viewpoint. (
  • Since the first DNA polymerase was discovered in 1958 , seven families of DNA polymerase have been described (A, B, C, D, X, Y and RT), each with different biochemical properties. (
  • These newly identified independent DNA polymerases are encoded by a new group of mobile genetic elements discovered by Mart Krupovic in bacteria and given the name "pipolins", based on the expression primer-independent polymerase. (
  • Primer-Independent DNA Synthesis by a Family B DNA Polymerase from Self-Replicating Mobile Genetic Elements, Cell reports , November 7, 2017. (
  • The Polymerase Chain Reaction, or PCR, is a basic method used in molecular biology to produce copies of a small target region of DNA in a sample. (
  • It has been shown that the cell cycle is deregulated in preimplantation mouse embryos treated with DPH by altering the expression of cyclin A. Even more, the DNA polymerase  (Pol ) protein catalytic subunit has been shown to have a decreased expression contributing to delayed DNA synthesis in 2-cell mouse embryos. (
  • The HotStarTaq Plus DNA Polymerase is intended for molecular biology applications. (
  • The polymerase combines the high specificity, sensitivity, and minimal optimization of HotStarTaq DNA Polymerase, with a fast 5-minute activation time. (
  • Each lot of HotStarTaq Plus DNA Polymerase is subjected to a comprehensive range of quality control tests, including a stringent PCR specificity and reproducibility assay in which low-copy targets are amplified. (
  • HotStarTaq Plus DNA Polymerase outperformed kits tested from other suppliers and ensures high specificity and superior performance in hot-start PCR (see figures " Highest specificity. PCR was performed with HotStarTaq Plus DNA Polymerase, HotStarTaq DNA Polymerase, and Taq DNA Polymerase from QIAGEN, and 3 hot-start PCR enzymes from the indicated suppliers. Parallel reactions were performed following the suppliers' recommendations, using 50 ng human genomic DNA. A 1.5 kb fragment of the human CFTR gene was amplified in 35 PCR cycles. M : markers. "> Highest specificity " and " Higher specificity with different primer-template systems. Higher specificity with different primer-template systems. "> Higher specificity with different primer-template systems ", and table). (
  • HotStarTaq Plus DNA Polymerase provides the unrivaled performance of HotStarTaq DNA Polymerase with a shortened activation time of just 5 minutes. (
  • HotStarTaq Plus DNA Polymerase, a modified form of QIAGEN Taq DNA Polymerase, is supplied in an inactive state that has no polymerase activity at ambient temperatures. (
  • This assay uses HPV L1 consensus polymerase chain reaction (PCR) with biotinylated PGMY09/11 primer sets and β-globin as an internal control for sample amplification. (
  • One novel strategy to enrich rare mutations at low variant allele fractions (VAFs) with quantitative polymerase chain reaction (qPCR) and Sanger sequencing was contrived by introducing artificial hairpins into amplicons to compete with primers, coined as the hairpin competition amplification (HCA) system. (
  • HCA differs from the previously reported work in which hairpins are formed to inhibit extension of wild-type DNA using 5-exonuclease-negative polymerase, where the readout is dependent on melting curve analysis after asymmetric PCR. (
  • Plasmid DNA extraction was carried out on twenty-nine cefoxitin-resistant selected isolates using the Kado and Lin method, while genotypic detection of plasmid-mediated AmpC gene was carried out by the polymerase chain reaction (PCR) assay. (
  • This assay uses Roche Linear Array HPV Genotyping test that is based on HPV L1 consensus polymerase chain reaction (PCR) with biotinylated PGMY09/11 primer sets. (
  • These primers amplify approximately 600 basepairs of the ITS1-5.8S-ITS2 region of the ribosomal cistron. (
  • These primers amplify approximately 620 basepairs of the 28S region of the ribosomal cistron. (
  • These primers amplify approximately 717 bp of the coding region of the EF-1α gene. (
  • These primers amplify approximately 495 bp of exons and introns at the 5' end of the β-tubulin gene. (
  • These primers amplify a section of the intergenic spacer in the ribosomal cistron. (
  • Development of primer sets designed for use with the PCR to amplify conserved genes from filamentous ascomycetes. (
  • Excessive concentrations of forward and reverse primers can also cause formation of primer dimer when the primers anneal and amplify themselves independent of the target DNA. (
  • 7 Primers designed to amplify the partial groEL gene encoding heat-shock protein of Anaplasma phagocytophilum EphplgroELF (5′-ATGGTATGCAGTTTGATCGC-3′) and EphplgroELR (5′-TCTACTCTGTCTTTGCGTTC-3′) were used and expected to yield a 625-bp product for Anaplasma phagocytophilum and for Anaplasma platys , respectively. (
  • The possibility of copying DNA without the need for a primer may be useful for DNA amplification, and the simplicity of this new replication mechanism could have a number of applications. (
  • There is a special set of ITS primers specifically for amplification of the ITS region of dermatophytes, especially Trichophyton (Gräser, 2000). (
  • The most important aspect of the target DNA to consider is the total number of copies in the reaction available for amplification. (
  • The target DNA provides the initial template for the amplification of the first set of products amplified and continues to provide the template for the remaining cycles. (
  • As PCR products are generated, they also provide copies of the target DNA used as a template for amplification. (
  • The Digene hc2 HPV DNA Test using Hybrid Capture 2 technology is a nucleic acid hybridization microplate assay with signal amplification. (
  • In dPCR, extracted DNA is subjected to get amplification in nanopartitions using target gene primers, the EvaGreen master mix, or fluorescently labeled targeted gene-specific probes. (
  • Using species-specific primers and TaqMan probes, we established a real-time quantitative PCR system for amplification of a fragment in the 658-bp cytochrome oxidase subunit I (COI) region from JBS specimens. (
  • It also includes biotinylated β-globin primers as an internal control for sample amplification. (
  • Results of the Digene hc2 HPV DNA Test and HPV typing based on the Roche prototype line blot assay have been previously released. (
  • The DNA extracts are used in the Digene hc2 HPV DNA Test. (
  • Although the ITS primers are universal for fungi, the D1D2 region of the large ribosomal subunit has better discrimination for yeasts, with primers NL-1 and NL-4. (
  • Over a decade ago, DNA barcoding was proposed as a fast, cost-efficient and simple taxonomic method based on the use of a unique, short and standardized gene region (cytochrome c oxidase 1, COI, for animals) for identifying specimens and expediting discovery of putative new species [ 1 ]. (
  • In profiling gene status from 12 lung cancer ctDNA samples and 16 thyroid cancer FNA DNA samples, HCA demonstrated a 100% concordance rate compared to ddPCR and commercial ARMS kit. (
  • Mutations in exons 5-8 of the p53 tumor suppressor gene were determined by direct DNA sequence analysis. (
  • The primer mix amplifies essentially all HPV types found in the genital tract along with the human β-globin gene. (
  • However, the proper primers for Pol  were designed, and the concentrations of MgSO4 and the primers were both optimized at 2mM and 0.125mM respectably. (
  • Therefore, bacterial genomic DNA will have far more copies of the target in a 50 ng sample than human DNA. (
  • If the bands from either of ''ara'' or ''sac'' show up at the expected bps against the DNA ladder, this proves that the genomic DNA is sucessfully extracted. (
  • The aim of this experiment was to extract genomic DNA from Bacillus subtilis strain ATCC 6633. (
  • Genomic DNA extracted from paraffin-embedded tissue with the QIAamp DNA-formalin-fixed, paraffin-embedded tissue procedure was amplified by using the primer sets DiBu-F(5′-GCTAGATATGCTACCAACAAAA-3′)/ITS1 R(5′-CTCAATGCGTCTGCAATTCGC-3′) and BuF2-(5-CATTTATGCTAGATATGCTACCAAC-3′)/ITS1-R. The products were fractionated on 2% agarose gel and stained with ethidium bromide. (
  • Tick samples (3-5 ticks) were frozen and mashed by liquid nitrogen and then deoxyribonucleic acid (DNA) was extracted using the G-spin genomic DNA extraction kit (iNrRON Biotechnology Inc., Republic of Korea). (
  • Mart Krupovic from the Institut Pasteur and Modesto Redrejo from the Autonomous University of Madrid, together with their teams, have discovered a group of family B DNA polymerases that can directly add a first nucleotide opposite its complementary nucleotide to kick-start the synthesis process. (
  • However, more copies of the target DNA will reduce specificity of the PCR reaction and likely produce a greater number of false products. (
  • Too much primer reduces specificity and this will allow primers to anneal in regions of the template that are not the target region. (
  • Ultramer DNA Oligonucleotides are generated by proprietary synthesis methods that deliver high-quality oligos up to 200 bases. (
  • IDT proprietary DNA synthesis equipment permits rapid, high-quality synthesis of nucleic acids. (
  • Along with this refined synthesis cycle, Ultramer DNA Oligos use a solid support that is specifically designed to synthesize low-yield, high‑quality oligos up to 200 bases in length. (
  • All previously known DNA polymerases needed a primer, which could come either from another protein or from a pre-existing DNA or RNA fragment. (
  • In some cases, these primers may not provide sufficient identification, and a protein coding region may be required. (
  • Furthermore, the DNA methyltransferase inhibitor 5-AZA-dC suppressed cell viability , cell number in S-phase, ALP activity and osteogenesis-related protein levels in osteoblasts treated with low doses of NaF. (
  • O6-Methylguanine-DNA methyltransferase (MGMT) is a ubiquitous DNA repair protein that can correct the mismatch of O6 alkyl guanine and directly reverse DNA damage, which plays a key role in the early repair process of DNA damage [ 7 ]. (
  • This protein probably interacts with specific regions of DNA and with other proteins to turn genes on or off. (
  • The vaginal swab is extracted to obtain DNA. (
  • it does not describe the procedure for purification of fungal DNA. (
  • Trichophyton species DNA is amplified very poorly by the ITS primer set used for most other molds. (
  • Sometimes, certain areas of the exome are incompletely sequenced, for example when DNA primers bind poorly. (
  • Primer concentration is one variable dependent on the total volume of the PCR reaction in order that sufficient copies of the primer find the target annealing sites. (
  • This document describes some of the target genes and primers that can be used for DNA sequence-based identification of fungi and the PCR conditions with which to use those primers. (
  • Other primer sets have been used for other genes, but those described below are the most consistently available in databases for the identification of yeasts and molds that are most likely to be identified in a clinical microbiology laboratory. (
  • Effects of fluoride on the proliferation and activation of osteoblasts by regulating methylation of the DNA repair genes MGMT and MLH1. (
  • NaF treatment led to methylation of the DNA repair genes MGMT and MLH1 in osteoblasts, resulting in cell proliferation and activation and causing the development of skeletal fluorosis. (
  • A team from the Institut Pasteur, working with a Spanish team from the Autonomous University of Madrid, has discovered a new group of DNA polymerases that can start DNA replication without the help of other proteins. (
  • DNA polymerases are enzymes in cells or viruses which are involved in the process of DNA replication. (
  • These results were confirmed by direct DNA sequence analysis. (
  • The total number of cycles for PCR should be reduced when higher concentrations of target DNA are in the reaction. (
  • Our high quality reagents are available for every workflow, including popular DNA assembly methods such as NEBuilder ® HiFi DNA Assembly and NEBridge ® Golden Gate Assembly . (
  • However, poor DNA quality and quantity are the major demerits of conventional PCR and real-time quantitative PCR (qPCR), as herbal products and spices are highly enriched in secondary metabolites such as polyphenolic compounds. (
  • We use a relatively simple calculation to dilute primers to a final concentration of 10 uM as shown starting with the primary primer concentration of 1 micro-grams (ug)/ micro-liter (ul). (
  • A crucial premise of DNA barcoding is that genetic variation within species (intra-specific) is lower than among species (inter-specific) [ 1 - 3 ], i.e., that a 'barcoding gap' exists [ 4 ] which allows unknown specimens to be identified as an existing species or flagged as a putative new species. (
  • It uses chemiluminescence for the qualitative detection of eighteen types of human papillomavirus (HPV) DNA in cervical specimens. (
  • Specimens containing the target DNA hybridize with the HR or LR HPV RNA probe cocktail. (
  • This study continues the analysis of the DNA Pol  subunit by beginning to examine the effect DPH has on the expression of Pol  mRNA. (
  • For identification of Fusarium isolates within species complexes, the EF-1α primers should be used (O'Donnell, 2009). (
  • For identification of Scedosporium , Aspergillus , and Penicillium isolates within species complexes, the β-tubulin primers should be used (Glass, 1995). (
  • DNA is being extracted from a world-wide collection of 83 isolates. (
  • There are several factors to consider when optimizing PCR such as total copies of target DNA, primer concentration, MgCl2 and deoxynucleotides, or dNTPs. (
  • In this article we will focus on two variables, the number of copies of the target DNA and primer concentration. (
  • Excessive primer concentration is perhaps one important factor that often causes generation of false products in a PCR. (
  • The final judgment on primer concentration will be viewed after products are electrophoresed on an agarose gel in order to show the number of products amplified. (
  • We observed an increase in fluorescence signal as the concentration of target DNA increased in PN-CP blended formulations (mock controls). (
  • The template DNA isolated from bacteria may consist of only a 2 million-base genome whereas the human genome has 3 billion bases. (
  • single-stranded DNA oligos specifically built for your homology-directed repair (HDR) experiments. (
  • But they all have one thing in common: they can only add new nucleotides (the building blocks of DNA) to a strand of DNA or RNA if they have a primer and an existing strand of complementary DNA to use as a template. (
  • These polymerases do not need the help of a primer to add new nucleotides. (
  • The strong bonds between those nucleotides can interfere with sequencing and primer binding due to their high melting temperature. (
  • The final diluted sample of target DNA is better diluted in water rather than buffer because buffers can interfere with difficult PCR amplifications. (
  • Primers are used extensively in genetic and molecular biology techniques. (
  • DNA extraction of Bacillus subtilis for materials required and protocol. (
  • We successfully DNA-barcoded 380 individuals of all 31 presently recognized species. (
  • Using species-specific primers, we successfully demonstrated the use of dPCR in the authentication of medicinal botanicals. (
  • By synergizing temporal optimization of reagent delivery, reagent composition, and integration mechanism, we achieve targeted integration of large DNA cargo at efficiencies nearing those of viral vector platforms with minimal toxicity. (
  • We then adapted a method and pipeline for COI metabarcoding using generalist primers that target the eukaryote diversity present. (
  • However, genotyping low-copy tumor DNA (ctDNA) in patients is challenging due to abundant wild DNA backgrounds. (
  • The forward and reverse primer must have an exact base match with the beginning and end of the target region. (
  • The amount of forward and reverse primer should be limited to reduce potential false priming. (
  • ara'' and ''sac'' forward and reverse primers are two tests, which will be used in PCR. (
  • Whether you are performing your first cloning experiment or constructing multi-fragment DNA assemblies, NEB ® has the solution for you. (
  • The copies of DNA produced by PCR provide researchers with sufficient copies for other applications in research including automated Sanger sequencing. (
  • Therefore, it is important that sufficient copies of the original target DNA are present in the reaction. (
  • In general, for unknown molds, the ITS region of the rDNA is used as the primary target with primers ITS-1 and ITS-4 as the most general primer set. (
  • Generating copies of a target DNA region using PCR applications is not as sensitive to the quality of the template DNA when compared to Sanger sequencing. (
  • Too many copies of the original target can lead to generation of false products early in PCR that also act as template DNA. (
  • PCR conditions generally recommend 10E4 to 10E5 copies of the target DNA in the reaction independent of the total volume. (
  • The results of excessive primers are often seen in unclean Sanger sequencing results because false products can be sequenced along with the desired target. (
  • The intensity of the light emitted denotes the presence or absence of target DNA in the specimen. (
  • The third-generation digital PCR (dPCR) technology is a highly sensitive, accurate, and reliable method to detect target DNA molecules as it is less affected by PCR inhibiting secondary metabolites due to nanopartitions. (
  • Note: We recommend all analysis of HPV DNA PCR be conducted using the Roche LA results (Data set name: L37SWR_C) in order to provide the most accurate longitudinal information on HPV detection and typing by PCR. (
  • Detection and typing of HPV DNA in vaginal swabs (in conjunction with testing of NHANES sera for HPV antibody) will allow evaluation of trends in prevalence of type-specific HPV infection by age, sexual behavior, and race/ethnicity. (
  • Here, we describe the detection of Carica papaya (CP) adulteration in Piper nigrum (PN) products using species-specific primers. (
  • Detection of the IS Ecp1B sequence munity, mostly in hospitals and often in ité de l'Antibiogramme de la Société was performed by PCR using primers intensive care units (ICUs) [3]. (
  • Methodology: This was a hospital-based, analytical cross-sectional study carried out on 226 symptomatic women wherein cervico-vaginal samples were obtained during gynaecological examination for Pap smears, HPV-DNA and genotype detection with linear array HPV strip, conducted from November 2019 to January 2021. (
  • Due to the significance of the Socotra Archipelago (a UNESCO Natural World Heritage site and a biodiversity hotspot) and the conservation interest of its reptile fauna (94% endemics), we performed the most comprehensive DNA barcoding study on an island group to date to test its applicability to specimen identification and species discovery. (
  • The influence imposed by artificial hairpins on primer-binding in a high-temperature PCR system was investigated for the first time in this work, paving the way for the optimization of HCA. (
  • The resultant RNA:DNA hybrids are captured onto the surface of a microplate well coated with antibodies specific for RNA:DNA hybrids. (
  • Immobilized hybrids are then reacted with alkaline phosphatase conjugated antibodies specific for the RNA:DNA hybrids, and detected with a chemiluminescent substrate. (
  • Authentication of herbal products and spices is experiencing a resurgence using DNA-based molecular tools, mainly species-specific assays and DNA barcoding. (
  • Herein we report a molecular quantification-based method for differentiation of fresh and dried JBS by determining the copy number of a specific DNA marker in the samples. (
  • The internal transcribed spacer (ITS) 1 PCR product (182 bp) was automatically sequenced by using the same primers used for PCR. (
  • Functions of Gtf2i and Gtf2ird1 in the developing brain: transcription, DNA binding and long-term behavioral consequences. (
  • Universal primer sets exist, but they often do not have enough discriminatory power to identify species, or they do not have the discriminatory power to identify species within a species complex, often giving 100% match to multiple species. (
  • For human DNA 10E5 will require over 300 nanograms of DNA, a one million fold difference. (
  • Design and test primers for this sequence using Primer-BLAST. (
  • With apologies to the incredible menagerie of design folk out there, this primer primarily focuses on the design denizens and design practices that surround software development. (
  • For the forward primer there are two options. (
  • Paquis-Flucklinger and colleagues reported that their myopathy patients had disorganized mitochondria with fragmented DNA. (
  • justifiant de ce fait une meilleure prise en charge de ces patients. (
  • La présente étude détermine la prévalence de l'infection par le virus de l'hépatite C en en determinant les génotypes ainsi que les facteurs y associés dans ce groupe de patients. (
  • Please note, the protocol is generic as all 38 second round primer pairs require the same master mix. (
  • Un test d'amplification en chaîne par polymérase (PCR) multiplex a également été mis au point pour identifier les isolats, et il s'est avéré que cette autre solution constituait un test diagnostique rapide, sensible et précis. (
  • Please note, the protocol is generic as all 38 primer pairs require the same master mix (see Appendix A). For each SARS-CoV-2 sample to be sequenced, 38 individual PCR reactions are required. (
  • In January 2007, our office released the first edition of California's Criminal Justice System: A Primer to provide the public, media, and policymakers some basic information on the state's criminal justice system, caseloads, costs, trends, and outcomes. (