A nucleic acid hybridization assay employing an immobilized or immobilizable polynucleotide probe selected to form DNA.RNA or RNA.RNA hybrids with the particular polynucleotide sequence to be determined. Resulting hybrids are detected by binding of an antibody reagent, preferably labeled with a detectable chemical group, selective for binding the hybrids in the presence of the single stranded sample and probe nucleic acids. No immobilization or labeling of sample nucleic acids is necessary and hybridization can be performed entirely in solution.
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Definition of Nucleic acid hybridization with photos and pictures, translations, sample usage, and additional links for more information.
Nucleic acid hybridization has been used in an attempt to determine the fraction of spleen DNA which is homologous to immunoglobulin messenger RNA (mRNA). Microsomes prepared from the mouse myeloma MOPC 104E and from spleens of mice hyperimmunized with Salmonella typhi were used as a source of RNA rich in immunoglobulin mRNA. It was shown that RNA extracted from microsomes contained a very limited number of RNA species and was essentially uncontaminated by other cytoplasmic and nuclear RNAs. Hyperimmune spleen DNA was hybridized with radiolabeled RNA from spleen and myeloma microsomes in the presence of unlabeled liver RNA. It was assumed that liver RNA should compete for the binding of most or all microsomal RNA species other than immunoglobulin mRNA. The amount of presumptive immunoglobulin mRNA bound indicated that spleen cells contain about 6 to 14 × 103 sequences per haploid genome for immunoglobulins of the size of the variable region of immunoglobulin polypeptide chains. The data favor ...
Background Optical imaging (OI) techniques such as bioluminescence and fluorescence imaging have been widely used to track diseases in a non-invasive manner within living subjects. These techniques generally require bioluminescent and fluorescent probes. Here we demonstrate the feasibility of using radioactive probes for in vivo molecular OI. Methodology/Principal Findings By taking the advantages of low energy window of light (1.2-3.1 eV, 400-1000 nm) resulting from radiation, radionuclides that emit charged particles such as β+ and β− can be successfully imaged with an OI instrument. In vivo optical images can be obtained for several radioactive probes including 2-deoxy-2-[18F]fluoro-D-glucose ([18F]FDG), Na18F, Na131I, 90YCl3 and a 90Y labeled peptide that specifically target tumors. Conclusions/Significance These studies demonstrate generalizability of radioactive OI technique. It provides a new molecular imaging strategy and will likely have significant impact on both small animal and
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Single-molecule measurements of DNA hybridization kinetics are mostly performed on a surface or inside a trap. Here we demonstrate a time-resolved, 3D single-molecule tracking (3D-SMT) method that allows us to follow a freely diffusing ssDNA molecule in solution for hundreds of milliseconds or even seconds a
Lesions in genetic sequences, for example, the sickle cell anemia mutation in the beta globin gene, are detected by means of interactive labels in a nucleic acid hybridization assay. A signal is gener
The Suppression Subtractive Hybridization (SSH) method has previously been used to generate tissue-specific cDNA libraries and to compare gene expression in different types of tissues. We propose that the SSH method can be modified to serve as a mutation detection method. This modified SSH method would scan the entire genome for sequence disruptions in DNA. Two plasmids have been chosen to serve as a model system. One plasmid is a PGL-2 plasmid, and the second plasmid is a PGL-2 plasmid that contains a 935 base pair (bp) segment of the G6PDH gene. The goal of this project is to detect the 935 bp insert in the PGL-2 plasmid. The purpose of the plasmid model system is to demonstrate that the modified SSH method is feasible, and successful as a mutation detection method.
complementary nucleotide sequences in a hybridization process. Tools in molecular biology relying on such a hybridization process include the polymerase chain reaction (PCR; and all methods based thereon), subtractive hybridisation, random primer extension, huclease S1 mapping, primer extension, reverse transcription, cDNA synthesis; differential display of RNAs, and DMA sequence determination, Northern blotting (RNA blotting), Southern blotting (DNA blotting). The hybridisation process can also occur with one of the complementary nucleic acids immobilised to a matrix such as magnetic beads, Sepharose beads or any other resin; Tools in molecular biology relying on such a process include the isolation of poly (A+) mRNA. The hybridisation process can furthermore occur with one of the complementary nucleic acids immobilised to a solid support such as a nitro -cellulose or nylon membrane or immobilised by e.g. photolithography to, for example, a siliceous glass support (the latter known as nucleic ...
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TY - JOUR. T1 - Novel differential gene expression in human cirrhosis detected by suppression subtractive hybridization. AU - Shackel, N. AU - McGuinness, Peter. AU - Abbott, Catherine. AU - Gorrell, M. AU - McCaughan, Geoffrey. PY - 2003. Y1 - 2003. M3 - Article. VL - 38. SP - 577. EP - 588. JO - Hepatology. JF - Hepatology. SN - 0270-9139. IS - 3. ER - ...
Nucleic acid fragments have been obtained from the genome of mycobacteria, and applications of the nucleic acid fragments in the diagnosis of mycobacterial infections are described. More particularly, the present invention concerns an isolated polynucleotide of the formula: 5-(SEQ ID NO: 1)-(formula III)-(SEQ ID NO: 2)-3 where formula III represents a polynucleotide containing nucleotides 343-1152 of SEQ ID NO: 3. Primers and probes based on the isolated polynucleotide, DNA complementary to any of the polynucleotides, primers or probes, a method of detecting and identifying at least one species or group of mycobacteria, and a kit, box, or coordinated set for conducting the method are also described.
Notice that A pairs with T and G pairs with C when a DNA strand hybridizes with another DNA strand. An RNA molecule can also form a base-paired DNA-RNA duplex molecule with a DNA that has complementary base pairing. The most common source of DNA complementary to an mRNA is the DNA coding strand that was the template for synthesis of the RNA. In DNA-RNA hybrid formation, G base pairs with C, A of the RNA pairs with T of the DNA, and U or the RNA pairs with A of the DNA. The DNA-RNA hybridization reaction is illustrated below: ...
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Looking for hybridization probe? Find out information about hybridization probe. A small molecule of deoxyribonucleic acid or ribonucleic acid that is radioactively labeled and used to identify complementary nucleic acid sequences by... Explanation of hybridization probe
A signal amplification method for detecting a target nucleic acid analyte having a homopolymeric region and a target sequence includes steps of: contacting an analyte under hybridizing conditions with a multiplicity of reporter probes, each reporter probe including a signal region and an oligonucleotide sequence which is complementary to and capable of forming a stable hybrid with the analyte homopolymeric region to form an analyte:reporter probe hybrid; and forming an analyte:capture probe hybrid by contacting the analyte target sequence with a capture probe under hybridizing conditions. The analyte:reporter probe hybrid may formed prior to contacting the analyte target sequence with the capture probe, so the result of contacting the analyte target sequence with the capture probe results in formation of an analyte:reporter probe:capture probe complex. The analyte:capture probe hybrid may be immobilized on a solid generally planar surface in an array format. Multiple reporter probes may form triple
The improvement of existing serological techniques, development of monoclonal antibody technology and the development of new serological approaches are all working together to provide new tools for the detection of disease-causing organisms in fish and crustaceans. Following the introduction of nucleic acid hybridization technique and PCR, it was recognized that the methods offered a sensitive approach to the detection and identification of specific microorganisms as in the case of a bacterial or viral infection in a variety of sample types. Potentially, a characteristic DNA sequence from a single virus particle or cell of a particular organism can be amplified to detectable levels within a short period of time. Conventional diagnostic methods that involve the culture of microorganisms can take days or weeks to complete or very tedious to perform. PCR offers a rapid, very sensitive, very specific and simple alternative. Further developments in immunodiagnostics and emerging technologies such as ...
Hello! Im using an in situ hybridization protocol utilizing DIG-labeled probes and alkaline phosphatase visualization to detect genomic sequences in tissue sections and blood spins. This is working excellently. Now Id like to do the same thing with FITC-labeled probes. The probe sequence is the same, the protocol is basically the same (before visualization steps, of course) - but I dont get any signal. I havent done FISH before but I have used fluorescently labeled antibodies, so there should be no problems in the basic handling of fluorescent components. The probe is commercially made and directly labeled with FITC. I have tried with two separately prepared lots, with no success. The in situ hybridization protocol is pretty standard. Any ideas? How about stability of the FITC label in denaturation, hybridization and washing steps? And how about sensitivity as compared with enzymatic detection? I get a strong signal with the AP system. For FISH, I have one FITC label per an oligonucleotide ...
i. To 3ug aRNA, add 3uL pdN15 (use PCR tubes).. 1. Or 2uL of pdN9 (5ug/uL).. 2. pdN15 = random pentadecamer.. 3. pdN9 = random nonamer.. 4. If you need to re-suspend a new tube of pdN15, use 350uL of water - Poly N (15-mer) 50 OD from Operon (Item # SP180-1) has a conc. of 31.7ug/OD, so total mass of 1585ug. Re-suspending in 350uL water gives a final conc. of 4.5ug/uL.. ii. Bring final volume to 19uL with water.. iii. Mix well and incubate at 70oC for 10min in PCR machine.. 1. Program 70 in Main folder (MJ PCR machine).. iv. Place on ice immediately and leave for at least 5min.. ...
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TY - JOUR. T1 - A hibridizáció és alkalmazása DNS-array rendszerekben. AU - Gyorffy, András. AU - Gyorffy, Balázs. AU - Molnár, Béla. AU - Tulassay, Zsolt. PY - 2005/12/1. Y1 - 2005/12/1. N2 - DNA hybridization arrays measure simultaneously the expression of several genes. First, a known DNA sequence (probe) is fixed on a firm basis. Then the complementer sequence (target sequence) is linked to it during the hybridization process. The target sequence extracted from biological samples is fluorescently, enzimatically or radioactivelly labeled before detection. Higher expression results in higher signal in the detection system. Unlabeled DNA strands can also be detected, as the electronical and optical characteristics of the DNA is altered after complementer hybridization. In this review we summarize the basics of hybridisation and its newest application area in the DNA array systems.. AB - DNA hybridization arrays measure simultaneously the expression of several genes. First, a known DNA ...
To exploit fully the potential of current sequencing technologies for population-based studies, one must enrich for loci from the human genome. Here we evaluate the hybridization-based approach by using oligonucleotide capture probes in solution to enrich for approximately 3.9 Mb of sequence target. We demonstrate that the tiling probe frequency is important for generating sequence data with high uniform coverage of targets. We obtained 93% sensitivity to detect SNPs, with a calling accuracy greater than 99%.
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A laboratory techniques|technique used to determine how similar two strands of single-stranded nucleic acids are to each other by putting them with a th...
SSC Hybridization Solution SSC solution is used for the hybridization of probes to nucleic acids. Contains: 6X SSC, 10% Dextran sulfate, 0.2% SDS, 5X Denhardts solution and sheared salmon sperm DNA... DNA/RNA Hybridization
2.2 Procedure. 1. Liquid hybridization was performed as the LHCD method, that is, isolated RNAs were hybridized in hybridization buffer (1 X Exo I buffer) with double-hairpin probe (aslo named as two-hairpin DNA probes) at 42°C (Li et al. 2012). The hybridization mixture was further incubated with FastDigest BamH I (Fermentas) for 1 h at 37°C according to the manufacturers instructions. 2. The mixture was digested with exonuclease I (NEB) for 5 min at 50°C and then for 1 h at 37°C. The short 50°C step must be performed to disrupt the short five-base-pair stem that remains on the capture oligo after Bam H I cleavage without disrupting hybridization of the miRNA to the capture oligo. 3. The digested mixture was used as the template for PCR, which was performed with KOD-Plus DNA polymerase according to the suppliers instructions. The PCR procedure was for 5 min at 94°C, then for 20 s at 94°C, for 20 s at 65°C, for 10 s at 68°C, X Cycles, and for 5 min at 68°C. The value of X was ...
2.2 Procedure. 1. Liquid hybridization was performed as the LHCD method, that is, isolated RNAs were hybridized in hybridization buffer (1 X Exo I buffer) with double-hairpin probe (aslo named as two-hairpin DNA probes) at 42°C (Li et al. 2012). The hybridization mixture was further incubated with FastDigest BamH I (Fermentas) for 1 h at 37°C according to the manufacturers instructions. 2. The mixture was digested with exonuclease I (NEB) for 5 min at 50°C and then for 1 h at 37°C. The short 50°C step must be performed to disrupt the short five-base-pair stem that remains on the capture oligo after Bam H I cleavage without disrupting hybridization of the miRNA to the capture oligo. 3. The digested mixture was used as the template for PCR, which was performed with KOD-Plus DNA polymerase according to the suppliers instructions. The PCR procedure was for 5 min at 94°C, then for 20 s at 94°C, for 20 s at 65°C, for 10 s at 68°C, X Cycles, and for 5 min at 68°C. The value of X was ...
The unsurpassed affinity, specificity, and rapid binding of yPNA make it the molecule of choice for demanding hybridization assays. Whether your lab is conducting basic research, developing tools or diagnostics, or involved in drug discovery and therapeutics, you should consider yPNA. The benefits of γPNA are being realized in fluorescence in situ hybridization (FISH) to measure chromosomal elements such as telomeres, to detect mRNA and miRNA, for antisense knockdown and gene silencing, as well as viral genotyping. Additionally, researchers have shown that γPNA can be used as to efficiently capture and pull out minute quantities of DNA and RNA from blood and other complex matrices for subsequent PCR and sequence analysis. There is great potential for γPNA in virtually any assay or application that relies on nucleic acid hybridization ...
DNA hybridization experiments demonstrated that the gene clusters encoding the F8 fimbriae (fei) as well as the type I fimbriae (pi/) exist in a single copy on the chromosome of E. coli 018:K5 strain 2980. In conjugation experiments with appropriate donors, the chromosomal site of these gene clusters was determined. The pil genes were mapped close to the gene clusters thr and Jeu controlling the biosynthesis of threonine and leucine, respectively. The fei genes were found to be located close to the galactose operon (gal) between the position 17 and 21 of the E. coli chromosomallinkage map ...
Sweet, R; Goodman, N; Cho, J; Ruprecht, R; Redfield, R; and Spiegelman, S, The presence of unique dna sequences after viral induction of leukemia in mice. (1974). Subject Strain Bibliography 1974. 32 ...
Advanced Cell Diagnostics team of in situ hybridization experts are ready to support you in getting superior results using our RNAscope® hybridization assays.
TY - JOUR. T1 - Evaluation of DNA sequencing ambiguities using tetramethyl-ammonium chloride hybridization conditions. AU - Connors, T. D.. AU - Burn, T. C.. AU - VanRaay, T.. AU - Germino, G. G.. AU - Klinger, K. W.. AU - Landes, G. M.. PY - 1997/6. Y1 - 1997/6. UR - http://www.scopus.com/inward/record.url?scp=0030914895&partnerID=8YFLogxK. UR - http://www.scopus.com/inward/citedby.url?scp=0030914895&partnerID=8YFLogxK. M3 - Article. C2 - 9187758. AN - SCOPUS:0030914895. VL - 22. SP - 1088. EP - 1090. JO - BioTechniques. JF - BioTechniques. SN - 0736-6205. IS - 6. ER - ...
At the time of publication of a paper by Hozumi and Tonegawa (1) in 1976, a controversy was raging about the number of germline variable (V) genes and how Ab diversity was generated (2). Protein sequencing had clearly shown that Abs were divided into two contiguous regions: a V region and a constant (C) region. Dreyer and Bennett (3) had earlier proposed the two gene-one polypeptide theory, which implied that the genes would somehow be joined at the DNA or RNA level. A plethora of papers were swimming in the literature, which tried to estimate the number of V genes based on liquid hybridization of RNA to DNA, all of which gave inconclusive results because the RNA was not pure, etc. It was widely accepted that there were a handful of C genes, and probably many handfuls of V genes. If so, how would a V gene find its way next to a C gene?. Out of the blue, Hozumi and Tonegawa (1) published a stunning paper that used newly discovered restriction enzymes to digest DNA. They showed by hybridization ...
The invention discloses and claims a signal amplification method for detecting a target nucleic acid analyte having a homopolymeric region and a target sequence. The method comprises (a) contacting an analyte under hybridizing conditions with a multiplicity of reporter probes, each probe including a signal region and an oligonucleotide sequence which is complementary to, and capable of forming a stable hybrid with the analyte homopolymeric region, whereby the hybridization of multiple reporter probes to the homopolymeric region provides for signal amplification; and (b) forming an analyte:capture probe hybrid by contacting the analyte target sequence with a capture probe under hybridizing conditions.
TSG-6 protein and functional derivatives thereof, DNA coding therefor, expression vehicles, such as a plasmids, and host cells transformed or transfected with the DNA molecule, and methods for producing the protein and the DNA are provided, as well as antibodies specific for the TSG-6 protein; a method for detecting the presence of TSG-6 protein in a biological sample; a method for detecting the presence of nucleic acid encoding a normal or mutant TSG-6 protein; a method for measuring induction of expression of TSG-6 in a cell using either nucleic acid hybridization or immunoassay; a method for identifying a compound capable of inducing the expression of TSG-6 in a cell; and a method for measuring the ability of a cell to respond to TNF.
Unlike optical trapping or electrical trapping, our method doesnt require perturbative force fields such as a strong electric field. It only uses gentle fluid flow to confine and manipulate particles. Were using it now to look at interactions between soft materials like vesicles or cells, such as how does the process of vesicle collision or adhesion happen? Our work tends to be focused on these fundamental questions, but these are materials that are used in foods, personal care products, detergents, and liquid fabric softeners, he said.. Looking ahead to new projects, Schroeder is excited about combining elements of biopolymers with synthetic polymers to look at the additive or new properties that arise. Specifically, theyre using elements from nucleic acid hybridization or base pairing to guide the structure of optoelectronic materials.. For the project in collaboration with Northwestern Universitys Michael Jewett and funded by the Department of Defenses Multidisciplinary University ...
Methods of detecting, probing, mapping and directed sequencing of target nucleic acids are provided using a guide RNA and a Cas9 protein. Methods for detecting the binding of the guide RNA/Cas9 complex to a target nucleic acid where the guide RNA includes a 3 tail sequence that can hybridize to a probe are provided. Methods for detecting the binding of the guide RNA/Cas9 complex to a target nucleic acid where the complex is physically detected are provided.
Northern assays require RNA (total- or poly(A)- selected) to be resolved on a agarose gel under denaturing conditions, transferred to a membrane and immobilized for subsequent hybridization. Various inconsistencies in the procedure, particularly the transfer set-up, can greatly effect the assays outcome. Here, we describe several technical points which can make any Northern analysis a success.
Yng Chen writes: , In Southern hybridization experiments how important is hybridization , temperature? What are the effects of rasing or lowering the hybridization , temperature? One generally hybridizes 15-20 C below the Tm to get favorable kinetics. So the exact temp. is not critical with respect to enforcing stringency. The only exception is if youre trying to do something at very low stringency, then you may need to lower the hybridization temp. below standard conditions. , How does one determine the proper washing conditions (assuming 100% , match in base paring). [for oligo probes] , If you get a weak signal and you know that you have a lot of DNA , on the membrane and that you have good specific activity on your , probe, how do you know if this is because your washing conditions , are too strigent or that the signal is non-specific. You start with a calculation, and then confirm by doing a temperature series against a cloned control target. If youre trying to discriminate a one base ...
Replacement vein for the Pitrainer. The Pitrainer sampling sleeve is intended for the hybrid formation of the blood sample or intravenous injection. It consists of a flexible sleeve simulating the skin under which a flexible armoured support is integrated for the protection of the wearer and which allows the insertion of a vein with bifurcation.
The ability of labeled DNA or RNA probes to bind with high affinity and specificity to complementary nucleic acid sequences forms the basis of nucleic acid hybridization assays. Immunoassays, on the other hand, exploit the strong and specific interaction between antigens and antibodies. One aspect of this dissertation deals with enhancement of the sensitivity of bioluminescence hybridization assays based on the photoprotein aequorin. This is achieved by introducing, enzymatically, multiple aequorin labels per DNA hybrid. The bound aequorin is determined by its characteristic bioluminescence. An 8-fold improvement in sensitivity is observed with the amplified assay as compared to an assay without the amplification step. Another aspect of this dissertation involves the development of two novel microtiter well-based DNA hybridization assays in which an expressible cDNA fragment encoding firefly luciferase serves as a reporter molecule. The reporter molecule undergoes in vitro expression through coupled
TY - JOUR. T1 - Poliovirus detection in water by cell culture and nucleic acid hybridization. AU - Enriquez, Carlos E.. AU - Abbaszadegan, Morteza. AU - Pepper, Ian L.. AU - Richardson, Kenneth J.. AU - Gerba, Charles P.. N1 - Copyright: Copyright 2015 Elsevier B.V., All rights reserved.. PY - 1993/7. Y1 - 1993/7. N2 - Nucleic acid hybridization has been used to detect viral nucleic acid in water. This type of assay, in contrast with tissue culture assays, may not distinguish between viable and non-viable viruses. We evaluated, by comparison with tissue culture infectivity assay (plaque forming method), the ability of the gene probe assay to detect viable poliovirus 1 (LSc) in well water, autoclaved well water, filter-sterilized well water and autoclaved phosphate buffered saline kept at 37 and 15°C for 75 days, and in dechlorinated tap water held at room temperature. A gradual decline in numbers of poliovirus was observed in all of the samples by cell culture assay. With the exception of ...
Comparative genomic hybridization analysis of adrenocortical tumors.: Comparative genomic hybridization (CGH) is a molecular cytogenetic technique that allows t
TY - JOUR. T1 - Comparison of nucleic acid hybridization and fluorometry for measurement of the relationship between RNA/DNA ratio and growth rate in a marine bacterium. AU - Kerkhof, L.. AU - Ward, B. B.. PY - 1993. Y1 - 1993. N2 - Continuous culture of Pseudomonas stutzeri Zobell, a marine denitrifying bacterium, was used to determine the relationship between growth rate and nucleic acid content. The trend of decreasing RNA content with decreasing growth rate, well known for enteric organisms, was found to occur in P. stutzeri Zobell as well, even at very long generation times such as those thought to occur in the oligotrophic ocean. When assayed by ethidium bromide fluorescence, the total RNA/DNA ratio was linear for generation times between 6 and 60 h. We also developed a 200-bp nucleic acid probe (with species- specific potential) for a portion of the 23S rRNA gene of P. stutzeri Zobell, which was used to quantify rRNA and rDNA by hybridization in the same continuous cultures. The rRNA/rDNA ...
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The accessibility and binding affinity of DNA are two key parameters affecting the hybridization efficiency in surface-based biosensor technologies. Better accessibility will result in a higher hybridization efficiency. Often, mixed ssDNA and mercaptohexanol monolayers are used to increase the hybridization efficiency and accessibility of surface-bound oligonucleotides to complementary target DNA. Here, no mercaptohexanol monolayer was used. We demonstrate by differential microcantilever deflection measurements at different pH that the hybridization efficiency peaks between pH 7.5 and 8.5. At low pH 4.5, hydration and electrostatic forces led to tensile surface stress, implying the reduced accessibility of the bound ssDNA probe for hybridization. In contrast, at high pH 8.5, the steric interaction between neighboring ssDNA strands was decreased by higher electrostatic repulsive forces, bending the microcantilever away from the gold surface to provide more space for the target DNA. Cantilever deflection
Nucleic acid sandwich hybridization assays are provided that incorporate one or a combination of background reduction steps. Those steps include use of a separate capture probe and separation from immobilized capture probes by cleavage and isolation. A very sensitive assay for RNA targets includes both of those steps, plus RNA binary probes, an RNA-directed RNA ligase and amplification by an RNA-directed RNA polymerase. Kits of reagents for performing assays according to this invention are also provided.
Looking for online definition of molecular hybridization in the Medical Dictionary? molecular hybridization explanation free. What is molecular hybridization? Meaning of molecular hybridization medical term. What does molecular hybridization mean?
17. S. Bhuckory, E. Hemmer, Y.-T. Wu, A. Yahia-Ammar, F. Vetrone, and N. Hildebrandt. Core or shell ? Er3+ FRET donors in upconversion nanoparticles. European Journal of Inorganic Chemistry 2017, 5186-5195.. 18. E. Porret, L. Sancey, A. Martín-Serrano, M. Montañez, R. Seemann, A. Yahia-Ammar, H. Okuno, F. Gomez, A. Ariza, N. Hildebrandt, J.-B. Fleury, J.-L. Coll, X. Le Guével. Hydrophobicity of Gold Nanoclusters Influences their Interactions with Biological Barriers. Chemistry of Materials 2017, 29 (17), 7497-7506.. 19. S. Díaz, G. Lasarte Aragones, S. Buckhout-White, X. Qiu, E. Oh, K. Susumu, J. Melinger, A. Huston, N. Hildebrandt, and I.L. Medintz. Bridging Lanthanide to Quantum Dot Energy Transfer with a Short Lifetime Organic Dye. The Journal of Physical Chemistry Letters 2017, 8 (10), 2182-2188.. 20. X. Qiu, J. Guo, Z. Jin, A. Petreto, I. L. Medintz, N. Hildebrandt. Multiplexed Nucleic Acid Hybridization Assays Using Single-FRET-Pair Distance-Tuning. Small 2017, 13, 1700332.. 21. M. ...
TY - JOUR. T1 - Suppression subtractive hybridization. AU - Ghorbel, Mohamed T. AU - Murphy, David. PY - 2011. Y1 - 2011. N2 - Comparing two RNA populations that differ from the effects of a single independent variable, such as a drug treatment or a specific genetic defect, can establish differences in the abundance of specific transcripts that vary in a population dependent manner. There are different methods for identifying differentially expressed genes. These methods include microarray, Serial Analysis of Gene Expression (SAGE), and quantitative Reverse-Transcriptase Polymerase Chain Reaction (qRT-PCR). Herein, the protocol describes an easy and cost-effective alternative that does not require prior knowledge of the transcriptomes under examination. It is specifically relevant when low levels of RNA starting material are available. This protocol describes the use of Switching Mechanism At RNA Termini Polymerase Chain Reaction (SMART-PCR) to amplify cDNA from small amounts of RNA. The ...
Summary of Facts and Submissions. I. European patent no. 0 497 784 was granted with 39 claims on the basis of European patent application 90913621.0 (published as WO 91/02817, referred to in this decision as the application as filed) and was opposed on the grounds of Article 100(a) EPC, for lack of novelty and inventive step (Articles 54 and 56 EPC), Article 100(b) EPC and Article 100(c) EPC.. II. Independent claims 1, 12 and 30 as granted read:. 1. Use of an internal standard for the quantitation of at least one target nucleic acid segment contained within a sample in an amplification method, said internal standard comprising on one strand a nucleic acid segment comprising a 5 sequence and a 3 sequence, which sequences provide an upstream primer hybridization site and the complement of a downstream primer hybridization site which are identical to an upstream primer hybridization site and the complement of a downstream primer hybridization site within said target nucleic acid segment, ...
Background: Genome-wide analysis of sequence divergence among species offers profound insights into the evolutionary processes that shape lineages. When full-genome sequencing is not feasible for a broad comparative study, we propose the use of array-based comparative genomic hybridization (aCGH) in order to identify orthologous genes with high sequence divergence. Here we discuss experimental design, statistical power, success rate, sources of variation and potential confounding factors. We used a spotted PCR product microarray platform from Drosophila melanogaster to assess sequence divergence on a gene-by-gene basis in three fully sequenced heterologous species (D. sechellia, D. simulans, and D. yakuba). Because complete genome assemblies are available for these species this study presents a powerful test for the use of aCGH as a tool to measure sequence divergence. Results: We found a consistent and linear relationship between hybridization ratio and sequence divergence of the sample to the ...
Biological RNA generally comprises secondary structure motifs which cause a problem for target RNA detection by isothermal amplification methods. The complexity of the secondary structures makes RNA targets inaccessible for probe hybridization, resulting in decreased sensitivity and selectivity. This is particularly important because the hybridization step of the isothermal amplification method re ...
Biological RNA generally comprises secondary structure motifs which cause a problem for target RNA detection by isothermal amplification methods. The complexity of the secondary structures makes RNA targets inaccessible for probe hybridization, resulting in decreased sensitivity and selectivity. This is particularly important because the hybridization step of the isothermal amplification method re ...
A fluorescently labeled nucleic acid having a hairpin structure between the fluorophore label and a point of attachment to a solid phase is useful as a probe to detect nucleic acid from a sample. The solid phase quenches the fluorophore label when the hairpin structure exists but this quenching is relieved by duplex formation between probe and a sample oligonucleotide. Probes for specific nucleic acid sequences can be immobilized as arrays on solid phase surfaces for detection of multiple nucleic acid sequences simultaneously from electrophoresis gels and from aqueous solutions. These probes and methods for their use can be combined with known solid phases, particularly those used for plasmon surface detection and electron transfer detection of nucleic acid. The probes can be washed and reused, and have other advantageous features over known probe methods.
An in situ molecular hybridization system which will detect retrovirus RNA in the cytoplasm of individual virus-infected cells has been developed. The technique was applied to cells infected with simian sarcoma-leukemia virus, where the virus-specific RNA was detected by hybridization to simian sarcoma-leukemia virus 3H-labeled complementary DNA. The system is useful for detecting viral RNA-containing cells in the presence of an excess of virus-negative cells and for determining which type of cell in a heterogenous population is expressing viral RNA.
TY - JOUR. T1 - Enhancement of surface plasmon resonance sensing for DNA hybridization using colloidal Au attached probe DNA. AU - Yamaguchi, Akira. AU - Juodkazis, Saulius. AU - Matsuo, Shigeki. AU - Misawa, Hiroaki. PY - 2002/2/5. Y1 - 2002/2/5. N2 - In this study, we demonstrate that the Au particle modified probe DNA monolayer can enhance the surface plasmon resonance (SPR) signal for measuring hybridization of unlabeled DNA molecules. The Au particles adsorbed on single stranded (ss)- and double stranded (ds)-DNA monolayers have different optical interaction with surface of Au thin film, and this difference induces the enhancement of the SPR signal.. AB - In this study, we demonstrate that the Au particle modified probe DNA monolayer can enhance the surface plasmon resonance (SPR) signal for measuring hybridization of unlabeled DNA molecules. The Au particles adsorbed on single stranded (ss)- and double stranded (ds)-DNA monolayers have different optical interaction with surface of Au thin ...
TY - JOUR. T1 - Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. AU - Melton, D. A.. AU - Krieg, P. A.. AU - Rebagliati, M. R.. AU - Maniatis, T.. AU - Zinn, K.. AU - Green, M. R.. N1 - Funding Information: ACKNOWLEDGEMENTS We are grateful to D. Ward and B. Mierendorf for sharing their unpublished results. We thank K. Breakey for help in preparing figures. P. K. acknowledges support from the Fogarty International Fellowship program. M.R. is grateful for support from an NSF Predoctoral Fellowship. This work was supported by a grant from the NIH to T.M. and grants from the NIH and the Chicago Community Trust/Searle Scholars Program to D.M. Copyright: Copyright 2010 Elsevier B.V., All rights reserved.. PY - 1984/9/25. Y1 - 1984/9/25. N2 - A simple and efficient method for synthesizing pure single stranded RNAs of virtually any structure is described. This in vitro transcription system is based on the ...
There is merit in each of these predictions. For example, profiling has re-emerged in clinical testing in the form of protein, tissue and nucleic acid arrays (e.g., cytokine profiles, array comparative genomic hybridization analysis) [49-52]. Computers and automation have played an increasingly important role in improving the efficiency and effectiveness of testing. More recently, pharmacogenetics, popularized by the slogan right patient, right drug, right time [53], has moved into mainstream testing (e.g., CYP2C9 for warfarin dosing) [54]. Since 1969, there have been many predictions and views of the future development of laboratory medicine and its sub-specialties. A summary of these predictions is provided in Table 1 [12-48]. A number of common themes and buzz-words can be identified in the prognostications such as nanotechnology, biosensors, microchips, genomics, and proteomics. These topics, together with the more specific predictions, are discussed in greater detail below. ...
TY - JOUR. T1 - A double labeling technique for performing immunocytochemistry and in situ hybridization in virus infected cell cultures and tissues. AU - Gendelman, Howard E.. AU - Moench, Thomas R.. AU - Narayan, Opendra. AU - Griffin, Diane E.. AU - Clements, Janice E.. PY - 1985/1/1. Y1 - 1985/1/1. N2 - This report describes a combined immunocytochemical and in situ hybridization procedure which allows visualization of cellular or viral antigens and viral RNA in the same cell. Cultures infected with visna or measles virus were fixed in periodate-lysine-paraformaldehyde-glutaraldehyde, stained by the avidin-biotin-peroxidase technique using antibodies to viral or cellular proteins and then incubated with radiolabeled specific DNA probes (in situ hybridization). The immunoperoxidase stain was preserved through the hybridization procedure. Nonspecific sticking of probes over peroxidase stained cells was prevented by incorporation of 0.1% Triton X-100 into the hybridization solution and the ...
We have confirmed and better defined the presence of a potentially important marker for malignant progression of BE. Furthermore, we have narrowed the area from 30 Mb, i.e., 7q33-q35 in our previous comparative genomic hybridization study (7) , to ∼2 Mb, i.e., the region between markers D7S2439 and D7S483. It implies the presence of a possible biomarker, which, in addition, has tumor suppressive activities. The 7q32.3-q36.1 region has not been reported frequently to be lost in human cancers. Thus far, it has been observed in gallbladder tumors, oral and oropharyngeal epithelial carcinomas, and leukemia (8, 9, 10) . In gallbladder tumors ,60% of allelic imbalance was seen for marker D7S798. Interestingly, it is located between D7S483 and D7S2465. We screened the critical area for known genes 7 with tumor suppressive potential and selected two possible candidates. Caspase 2 is known to stimulate apoptosis and is involved in shedding of intestinal epithelium (11) . Loss of these functions could ...
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Comparative genomic hybridization (CGH) is a modern genetic method which enables a genome‐wide survey of chromosomal imbalances. For each chromosome region, one obtains the information whether there is a loss or gain of genetic material, or whether t
Array hybridization devices and methods for their use are provided. The subject devices are characterized by having a substantially planar bottom surface, a cover, at least one fluid port and at least one adjustable spacing element for adjusting the spacing between an array and the bottom surface. In using the subject devices, an array is placed on the at least one adjustable spacing element in the chamber and the space between the array and the bottom surface is adjusted by moving the at least one adjustable spacing element. The adjusted array is contacted with at least one biological sample introduced into the chamber. The subject inventions find use in a variety of array-based applications, including nucleic acid array hybridizations.
|p>Decrease any non-specific hybridization of a probe to a substrate in Northern, Southern, or other nucleic acid hybridisation protocols. The Placentas used for obtaining the DNA come from healthy donors negative for the following: Hep-B & Hep C; HIV-1 and HIV-2; HTLV-1 and HTLV-1; Treponema Pallidum|/p>
The present invention includes polynucleotides encoding an MCP1 (e.g., a mature MCP1 polypeptide), an MCP1 multimer or a fusion thereof (e.g., fused to an in vivo half-life extending moiety (e.g., Ig)) (e.g., any of SEQ ID NOs: 1, 2, 8-12) as well as nucleic acids which hybridize to the polynucleotides. Preferably, the nucleic acids hybridize under low stringency conditions, more preferably under moderate stringency conditions and most preferably under high stringency conditions. A nucleic acid molecule is hybridizable to another nucleic acid molecule, such as a cDNA, genomic DNA, or RNA, when a single stranded form of the nucleic acid molecule can anneal to the other nucleic acid molecule under the appropriate conditions of temperature and solution ionic strength (see Sambrook, et a/., supra). The conditions of temperature and ionic strength determine the stringency of the hybridization. Typical low stringency hybridization conditions are 55°C, 5X SSC, 0.1% SDS, 0.25% milk, and no ...
Paraffin sections of human skeletal tissues were studied in order to identify cells responsible for production of types I, II, and III collagens by in situ hybridization. Northern hybridization and sequence information were used to select restriction fragments of cDNA clones for the corresponding mRNAs to obtain probes with a minimum of cross-hybridization. The specificity of the probes was proven in hybridizations to sections of developing fingers: osteoblasts and chondrocytes, known to produce only one type of fibrillar collagen each (I and II, respectively) were only recognized by the corresponding cDNA probes. Smooth connective tissues exhibited variable hybridization intensities with types I and III collagen cDNA probes. The technique was used to localize the activity of type II collagen production in the different zones of cartilage during the growth of long bones. Visual inspection and grain counting revealed the highest levels of pro alpha 1(II) collagen mRNAs in chondrocytes of the ...
This Decision Document has been prepared to explain the regulatory decision reached under the guidelines Dir94-08 Assessment Criteria for Determining Environmental Safety of Plants with Novel Traits and its companion document Dir94-09 The Biology of Brassica napus L. (Canola/Rapeseed), and the proposed guidelines Pro94-04 Guidelines for the Assessment of Plants with Novel Traits as Livestock Feed.
This Decision Document has been prepared to explain the regulatory decision reached under the guidelines Dir94-08 Assessment Criteria for Determining Environmental Safety of Plants with Novel Traits and its companion document Dir94-09 The Biology of Brassica napus L. (Canola/Rapeseed) and the guidelines Dir95-03 Guidelines for the Assessment of Livestock Feed from Plants with Novel Traits.
This Decision Document has been prepared to explain the regulatory decision reached under the guidelines Dir94-08 Assessment Criteria for Determining Environmental Safety of Plants with Novel Traits and its companion document Dir94-09 The Biology of Brassica napus L. (Canola/Rapeseed) and the guidelines Dir95-03 Guidelines for the Assessment of Livestock Feed from Plants with Novel Traits.
В немецком издательстве Springer Verlag вышла книга Fluorescence In Situ Hybridization (FISH). Application Guide (с годом издания 2017). Это второе издание сборника протоколов, который был выпущен в 2009 году. Три главы из этой книги написаны сотрудниками Отдела разнообразия и эволюции геномов ИМКБ или при их участии.. Yang F, Trifonov V, Ng BL, Kosyakova N, Carter NP. Generation of paint probes from flow-sorted and microdissected chromosomes. pp. 63-79. Trifonov VA, Vorobieva NV, Serdyukova NA, Rens W. FISH with and without COT1 DNA. pp. 123-132. Yang F, Graphodatsky AS. Animal probes and ZOO-FISH. pp. 395-415. This manual offers detailed protocols for fluorescence in situ hybridization (FISH) and comparative genomic hybridization approaches, which have been successfully used to study various aspects of ...
TY - JOUR. T1 - Drugging the R-loop interactome. T2 - RNA-DNA hybrid binding proteins as targets for cancer therapy. AU - Boros-Oláh, Beáta. AU - Dobos, Nikoletta. AU - Hornyák, Lilla. AU - Szabó, Zoltán. AU - Karányi, Zsolt. AU - Halmos, Gábor. AU - Roszik, Jason. AU - Székvölgyi, Lóránt. PY - 2019/12. Y1 - 2019/12. N2 - Unravelling the origin of genetic alterations from point mutations to chromosomal rearrangements was greatly enhanced by the discovery of RNA-DNA hybrids (R-loops) that behave as hotspots of genomic instability in a variety of organisms. Current models suggest that uncontrolled R-loops are a hazard to genome integrity, therefore, identifying proteins that are involved in recognising and signalling R-loop structures are of key importance. Herein we analysed key RNA-DNA hybrid binding proteins in humans taking advantage of large-scale gene expression, survival rate, and drug-sensitivity data from cancer genomics databases. We show that expression of RNA-DNA hybrid ...
TY - JOUR. T1 - Kinetics of spontaneous displacement of RNA from heteroduplexes by DNA. AU - Landgraf, Ralf. AU - Ramamurthi, Kumaran S.. AU - Sigman, David S.. PY - 1996/9/4. Y1 - 1996/9/4. N2 - We have used R-loop formation and direct hybridization techniques to analyze the kinetics by which RNA is displaced from a heteroduplex by DNA of identical sequence. Using random walk simulations we were able to calculate the step times for a single displacement reaction. For RNA with a GC content of 57-60% the data indicate an RNA exchange probability of 50.06%, which is indicative of a modest destabilization of the heteroduplex compared with a DNA duplex in the presence of magnesium. The average step time for the reversible exchange of a single nucleotide is 345.0 (± 1.3) ms/step. An acceleration of the displacement reaction was observed in the absence of magnesium. A comparison with step times for elongation shows that RNA displacement would not be rate limiting to transcription elongation under two ...
Rare DNA-sequence variants hold important clinical and biological information, but existing detection techniques are expensive, complex, allele-specific, or dont allow for significant multiplexing. Here, we report a temperature-robust polymerase-chain-reaction method, which we term blocker displacement amplification (BDA), that selectively amplifies all sequence variants, including single-nucleotide variants (SNVs), within a roughly 20-nucleotide window by 1,000-fold over wild-type sequences. This allows for easy detection and quantitation of hundreds of potential variants originally at ≤0.1% in allele frequency. BDA is compatible with inexpensive thermocycler instrumentation and employs a rationally designed competitive hybridization reaction to achieve comparable enrichment performance across annealing temperatures ranging from 56 °C to 64 °C. To show the sequence generality of BDA, we demonstrate enrichment of 156 SNVs and the reliable detection of single-digit copies. We also show that the BDA
Generation of bovine RPE/retina-subtracted cDNA library. Detailed description of the library will be published elsewhere (J. T. Chang, N. Della, C. Chew, S. Zhang, P. A. Campochiaro, and D. J. Zack, unpublished data). In brief, a library was constructed in Uni-ZAP XR (Strategene, La Jolla, CA) using cDNA that was generated from bovine RPE RNA; the library was in vivo excised and made single-stranded, hybridized in several rounds with an excess of biotinylated heart and liver RNA; the resulting RNA-DNA hybrids and unhybridized RNA were removed by phenol extraction after the addition of streptavidin; and the remaining unhybridized plasmid DNA was electroporated into MC1061 cells.. Fluorescent in situ hybridization. Fluorescentin situ hybridization (FISH) mapping was performed by standard methods (Lichter et al., 1990). Identical results were obtained with two independent but overlapping P1 clones. The clones were identified from high-density filters and were processed according to the suppliers ...
Advertisement Molecular Pathology. Microarray. Variations Tissue microarray. Author: Rodney E. Shackelford, DO, Ph.D. (see Reviewers page). Revised: 3 July 2010, last major update July 2010. Copyright: (c) 2008-2010, PathologyOutlines.com, Inc.. General. =========================================================================. ● Tissue microarray (TMA) significantly differs from the other techniques discussed in this chapter because the hybridization step involves antibody binding to one target protein, or nucleic acid hybridization to one target gene. ● TMA allows for the simultaneous analysis of protein expression in up to 500 different tissue samples. Advantages. =========================================================================. ● The relative level of a specific protein s expression can be compared on the same slide, allowing uniform analysis. ● A small amount of tissue is used per sample analyzed, allowing more analyses per same tissue volume and lowering the amount of ...
Fingerprint Dive into the research topics of Identification of underexpressed genes in early- and late-stage primary ovarian tumors by suppression subtraction hybridization. Together they form a unique fingerprint. ...
Hybrid organism is produced by interbreeding between two animals or plants of same or different taxa. This process of hybrid formation is hybridization.
A method of detecting a target nucleic acid A is disclosed, comprising hybridizing the target nucleic acid A with a probe nucleic acid B which contains a sequence B1 which base pairs with a part of the target nucleic acid A and a sequence B2, cleaving the hybridized probe nucleic acid B to produce a cleavage product B containing the sequence B2, hybridizing the cleavage product B with a template nucleic acid C containing a sequence C2 which base pairs with a part of the cleavage product B and a sequence C1 which does not hybridize with the sequence B1 of the probe nucleic acid B, extending the hybridized cleavage product B with an extension sequence B3 which is template-specific to a part of the sequence C1, hybridizing a probe D with the extension product, wherein the probe D contains a sequence D1 which base pairs with the extension sequence B3 and a sequence D2, and detecting any of the various products formed throughout the method. Products for performing the method are also disclosed.
Researchers have for the first time determined that hybridization between two bird species can give rise to several novel and fully functional hybrid genomic combinations. This could potentially be because hybrid species emerged through independent hybridisation events between the same parent species on different islands.
Fungi play important roles in the ecosystem processes such as nutrient cycling and degradation. However, the majority of fungi are unknown and difficult to isolate and culture. Due to the limitations of culture-based methods and traditional morphological observation, fungal ecologists turn to utilize molecular techniques such as nucleic acid hybridization,DNA sequencing,DNA fingerprinting techniques for the studies of species identification,population structure and community diversity. The present paper summarizes the theories of these molecular techniques and their applications in the study of fungal ecology.
Animals, Base Sequence, Brain Chemistry, Cell Line, Cerebellum, analysis, Cerebral Cortex, DNA, Female, Hippocampus, Hypothalamus, Nucleic Acid Hybridization, Poly A, isolation... ...