Cosmids: Plasmids containing at least one cos (cohesive-end site) of PHAGE LAMBDA. They are used as cloning vehicles.Cloning, Molecular: 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.Chromosomes, Artificial, Yeast: Chromosomes in which fragments of exogenous DNA ranging in length up to several hundred kilobase pairs have been cloned into yeast through ligation to vector sequences. These artificial chromosomes are used extensively in molecular biology for the construction of comprehensive genomic libraries of higher organisms.Restriction Mapping: Use of restriction endonucleases to analyze and generate a physical map of genomes, genes, or other segments of DNA.Chromosome Mapping: Any method used for determining the location of and relative distances between genes on a chromosome.Nucleic Acid Hybridization: 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)Genes, Bacterial: The functional hereditary units of BACTERIA.Gene Library: 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.Rhizobium: A genus of gram-negative, aerobic, rod-shaped bacteria that activate PLANT ROOT NODULATION in leguminous plants. Members of this genus are nitrogen-fixing and common soil inhabitants.Base Sequence: The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.Genomic Library: A form of GENE LIBRARY containing the complete DNA sequences present in the genome of a given organism. It contrasts with a cDNA library which contains only sequences utilized in protein coding (lacking introns).Plasmids: Extrachromosomal, usually CIRCULAR DNA molecules that are self-replicating and transferable from one organism to another. They are found in a variety of bacterial, archaeal, fungal, algal, and plant species. They are used in GENETIC ENGINEERING as CLONING VECTORS.Molecular Sequence Data: 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.Genetic Complementation Test: A test used to determine whether or not complementation (compensation in the form of dominance) will occur in a cell with a given mutant phenotype when another mutant genome, encoding the same mutant phenotype, is introduced into that cell.DNA Probes: 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.DNA Restriction Enzymes: Enzymes that are part of the restriction-modification systems. They catalyze the endonucleolytic cleavage of DNA sequences which lack the species-specific methylation pattern in the host cell's DNA. Cleavage yields random or specific double-stranded fragments with terminal 5'-phosphates. The function of restriction enzymes is to destroy any foreign DNA that invades the host cell. Most have been studied in bacterial systems, but a few have been found in eukaryotic organisms. They are also used as tools for the systematic dissection and mapping of chromosomes, in the determination of base sequences of DNAs, and have made it possible to splice and recombine genes from one organism into the genome of another. EC 3.21.1.Multigene Family: A set of genes descended by duplication and variation from some ancestral gene. Such genes may be clustered together on the same chromosome or dispersed on different chromosomes. Examples of multigene families include those that encode the hemoglobins, immunoglobulins, histocompatibility antigens, actins, tubulins, keratins, collagens, heat shock proteins, salivary glue proteins, chorion proteins, cuticle proteins, yolk proteins, and phaseolins, as well as histones, ribosomal RNA, and transfer RNA genes. The latter three are examples of reiterated genes, where hundreds of identical genes are present in a tandem array. (King & Stanfield, A Dictionary of Genetics, 4th ed)Chromosome Walking: A technique with which an unknown region of a chromosome can be explored. It is generally used to isolate a locus of interest for which no probe is available but that is known to be linked to a gene which has been identified and cloned. A fragment containing a known gene is selected and used as a probe to identify other overlapping fragments which contain the same gene. The nucleotide sequences of these fragments can then be characterized. This process continues for the length of the chromosome.WAGR Syndrome: A contiguous gene syndrome associated with hemizygous deletions of chromosome region 11p13. The condition is marked by the combination of WILMS TUMOR; ANIRIDIA; GENITOURINARY ABNORMALITIES; and INTELLECTUAL DISABILITY.DNA, Recombinant: Biologically active DNA which has been formed by the in vitro joining of segments of DNA from different sources. It includes the recombination joint or edge of a heteroduplex region where two recombining DNA molecules are connected.Hybrid Cells: Any cell, other than a ZYGOTE, that contains elements (such as NUCLEI and CYTOPLASM) from two or more different cells, usually produced by artificial CELL FUSION.In Situ Hybridization, Fluorescence: A type of IN SITU HYBRIDIZATION in which target sequences are stained with fluorescent dye so their location and size can be determined using fluorescence microscopy. This staining is sufficiently distinct that the hybridization signal can be seen both in metaphase spreads and in interphase nuclei.Blotting, Southern: 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.DNA, Bacterial: Deoxyribonucleic acid that makes up the genetic material of bacteria.Herpesvirus 3, Human: The type species of VARICELLOVIRUS causing CHICKENPOX (varicella) and HERPES ZOSTER (shingles) in humans.Genetic Markers: A phenotypically recognizable genetic trait which can be used to identify a genetic locus, a linkage group, or a recombination event.Genes: A category of nucleic acid sequences that function as units of heredity and which code for the basic instructions for the development, reproduction, and maintenance of organisms.Escherichia coli: 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.DNA Transposable Elements: Discrete segments of DNA which can excise and reintegrate to another site in the genome. Most are inactive, i.e., have not been found to exist outside the integrated state. DNA transposable elements include bacterial IS (insertion sequence) elements, Tn elements, the maize controlling elements Ac and Ds, Drosophila P, gypsy, and pogo elements, the human Tigger elements and the Tc and mariner elements which are found throughout the animal kingdom.Chromosomes, Human, Pair 3: A specific pair of human chromosomes in group A (CHROMOSOMES, HUMAN, 1-3) of the human chromosome classification.Deoxyribonuclease EcoRI: One of the Type II site-specific deoxyribonucleases (EC 3.1.21.4). It recognizes and cleaves the sequence G/AATTC at the slash. EcoRI is from E coliRY13. Several isoschizomers have been identified. EC 3.1.21.-.Chromosomes, Bacterial: Structures within the nucleus of bacterial cells consisting of or containing DNA, which carry genetic information essential to the cell.Chromosome Banding: Staining of bands, or chromosome segments, allowing the precise identification of individual chromosomes or parts of chromosomes. Applications include the determination of chromosome rearrangements in malformation syndromes and cancer, the chemistry of chromosome segments, chromosome changes during evolution, and, in conjunction with cell hybridization studies, chromosome mapping.Genetic Vectors: DNA molecules capable of autonomous replication within a host cell and into which other DNA sequences can be inserted and thus amplified. Many are derived from PLASMIDS; BACTERIOPHAGES; or VIRUSES. They are used for transporting foreign genes into recipient cells. Genetic vectors possess a functional replicator site and contain GENETIC MARKERS to facilitate their selective recognition.Mutagenesis, Insertional: Mutagenesis where the mutation is caused by the introduction of foreign DNA sequences into a gene or extragenic sequence. This may occur spontaneously in vivo or be experimentally induced in vivo or in vitro. Proviral DNA insertions into or adjacent to a cellular proto-oncogene can interrupt GENETIC TRANSLATION of the coding sequences or interfere with recognition of regulatory elements and cause unregulated expression of the proto-oncogene resulting in tumor formation.Deoxyribonuclease HindIII: One of the Type II site-specific deoxyribonucleases (EC 3.1.21.4). It recognizes and cleaves the sequence A/AGCTT at the slash. HindIII is from Haemophilus influenzae R(d). Numerous isoschizomers have been identified. EC 3.1.21.-.X Chromosome: The female sex chromosome, being the differential sex chromosome carried by half the male gametes and all female gametes in human and other male-heterogametic species.Genetic Linkage: The co-inheritance of two or more non-allelic GENES due to their being located more or less closely on the same CHROMOSOME.Ampicillin: Semi-synthetic derivative of penicillin that functions as an orally active broad-spectrum antibiotic.Time Factors: Elements of limited time intervals, contributing to particular results or situations.Ampicillin Resistance: Nonsusceptibility of a microbe to the action of ampicillin, a penicillin derivative that interferes with cell wall synthesis.Culture Media: Any liquid or solid preparation made specifically for the growth, storage, or transport of microorganisms or other types of cells. The variety of media that exist allow for the culturing of specific microorganisms and cell types, such as differential media, selective media, test media, and defined media. Solid media consist of liquid media that have been solidified with an agent such as AGAR or GELATIN.Smallpox Vaccine: A live VACCINIA VIRUS vaccine of calf lymph or chick embryo origin, used for immunization against smallpox. It is now recommended only for laboratory workers exposed to smallpox virus. Certain countries continue to vaccinate those in the military service. Complications that result from smallpox vaccination include vaccinia, secondary bacterial infections, and encephalomyelitis. (Dorland, 28th ed)Smallpox: An acute, highly contagious, often fatal infectious disease caused by an orthopoxvirus characterized by a biphasic febrile course and distinctive progressive skin eruptions. Vaccination has succeeded in eradicating smallpox worldwide. (Dorland, 28th ed)War: Hostile conflict between organized groups of people.Epilepsy: A disorder characterized by recurrent episodes of paroxysmal brain dysfunction due to a sudden, disorderly, and excessive neuronal discharge. Epilepsy classification systems are generally based upon: (1) clinical features of the seizure episodes (e.g., motor seizure), (2) etiology (e.g., post-traumatic), (3) anatomic site of seizure origin (e.g., frontal lobe seizure), (4) tendency to spread to other structures in the brain, and (5) temporal patterns (e.g., nocturnal epilepsy). (From Adams et al., Principles of Neurology, 6th ed, p313)Vaccinia virus: The type species of ORTHOPOXVIRUS, related to COWPOX VIRUS, but whose true origin is unknown. It has been used as a live vaccine against SMALLPOX. It is also used as a vector for inserting foreign DNA into animals. Rabbitpox virus is a subspecies of VACCINIA VIRUS.Directories as Topic: Lists of persons or organizations, systematically arranged, usually in alphabetic or classed order, giving address, affiliations, etc., for individuals, and giving address, officers, functions, and similar data for organizations. (ALA Glossary of Library and Information Science, 1983)Product Line Management: Management control systems for structuring health care delivery strategies around case types, as in DRGs, or specific clinical services.Biological Science Disciplines: All of the divisions of the natural sciences dealing with the various aspects of the phenomena of life and vital processes. The concept includes anatomy and physiology, biochemistry and biophysics, and the biology of animals, plants, and microorganisms. It should be differentiated from BIOLOGY, one of its subdivisions, concerned specifically with the origin and life processes of living organisms.Sensitivity and Specificity: 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)Biomedical Research: Research that involves the application of the natural sciences, especially biology and physiology, to medicine.DirectoryCatalogs as Topic: Ordered compilations of item descriptions and sufficient information to afford access to them.

Analysis of two cosmid clones from chromosome 4 of Drosophila melanogaster reveals two new genes amid an unusual arrangement of repeated sequences. (1/1067)

Chromosome 4 from Drosophila melanogaster has several unusual features that distinguish it from the other chromosomes. These include a diffuse appearance in salivary gland polytene chromosomes, an absence of recombination, and the variegated expression of P-element transgenes. As part of a larger project to understand these properties, we are assembling a physical map of this chromosome. Here we report the sequence of two cosmids representing approximately 5% of the polytenized region. Both cosmid clones contain numerous repeated DNA sequences, as identified by cross hybridization with labeled genomic DNA, BLAST searches, and dot matrix analysis, which are positioned between and within the transcribed sequences. The repetitive sequences include three copies of the mobile element Hoppel, one copy of the mobile element HB, and 18 DINE repeats. DINE is a novel, short repeated sequence dispersed throughout both cosmid sequences. One cosmid includes the previously described cubitus interruptus (ci) gene and two new genes: that a gene with a predicted amino acid sequence similar to ribosomal protein S3a which is consistent with the Minute(4)101 locus thought to be in the region, and a novel member of the protein family that includes plexin and met-hepatocyte growth factor receptor. The other cosmid contains only the two short 5'-most exons from the zinc-finger-homolog-2 (zfh-2) gene. This is the first extensive sequence analysis of noncoding DNA from chromosome 4. The distribution of the various repeats suggests its organization is similar to the beta-heterochromatic regions near the base of the major chromosome arms. Such a pattern may account for the diffuse banding of the polytene chromosome 4 and the variegation of many P-element transgenes on the chromosome.  (+info)

Nonmethylated transposable elements and methylated genes in a chordate genome. (2/1067)

The genome of the invertebrate chordate Ciona intestinalis was found to be a stable mosaic of methylated and nonmethylated domains. Multiple copies of an apparently active long terminal repeat retrotransposon and a long interspersed element are nonmethylated and a large fraction of abundant short interspersed elements are also methylation free. Genes, by contrast, are predominantly methylated. These data are incompatible with the genome defense model, which proposes that DNA methylation in animals is primarily targeted to endogenous transposable elements. Cytosine methylation in this urochordate may be preferentially directed to genes.  (+info)

Three receptor genes for plasminogen related growth factors in the genome of the puffer fish Fugu rubripes. (3/1067)

Plasminogen related growth factors (PRGFs) and their receptors play major roles in embryogenesis, tissue regeneration and neoplasia. In order to investigate the complexity and evolution of the PRGF receptor family we have cloned and sequenced three receptors for PRGFs in the teleost fish Fugu rubripes, a model vertebrate with a compact genome. One of the receptor genes isolated encodes the orthologue of mammalian MET, whilst the other two may represent Fugu rubripes orthologues of RON and SEA. This is the first time three PRGF receptors have been identified in a single species.  (+info)

Complete exon-intron organization of the mouse fibulin-1 gene and its comparison with the human fibulin-1 gene. (4/1067)

Fibulin-1 is a 90 kDa calcium-binding protein present in the extracellular matrix and in the blood. Two major variants, C and D, differ in their C-termini as well as the ability to bind the basement membrane protein nidogen. Here we characterized genomic clones encoding the mouse fibulin-1 gene, which contains 18 exons spanning at least 75 kb of DNA. The two variants are generated by alternative splicing of exons in the 3' end. By searching the database we identified most of the exons encoding the human fibulin-1 gene and showed that its exon-intron organization is similar to that of the mouse gene.  (+info)

Genetic variation of chlorella viruses: variable regions localized on the CVK2 genomic DNA. (5/1067)

A physical map of the Chlorella virus CVK2 genomic DNA has been constructed based on a cosmid contig covering the entire genomic region. By using Southern blot analysis with 22 gene probes, the gene arrangement along the genome was compared between CVK2 and PBCV-1, the prototypic member of Phycodnaviridae, whose genomic sequence is now available. The major rearrangements were (1) an insertion of a 20-kbp region around the left end of CVK2 DNA, (2) a duplication of the gene for major capsid protein in CVK2 DNA, (3) deletions/insertions of some open reading frames, and (4) divergence in the terminal inverted repeat sequences. Despite these changes, extensive colinearity was revealed between most of the genes along the CVK2 and PBCV-1 genomes. These data imply that the Chlorella virus genome has an overall high degree of genomic stability, encompassing specific islands of rearrangements.  (+info)

Comparative genomic analysis of the interferon/interleukin-10 receptor gene cluster. (6/1067)

Interferons and interleukin-10 are involved in key aspects of the host defence mechanisms. Human chromosome 21 harbors the interferon/interleukin-10 receptor gene cluster linked to the GART gene. This cluster includes both components of the interferon alpha/beta-receptor (IFNAR1 and IFNAR2) and the second components of the interferon gamma-receptor (IFNGR2) and of the IL-10 receptor (IL10R2). We report here the complete gene content of this GART-cytokine receptor gene cluster and the use of comparative genomic analysis to identify chicken IFNAR1, IFNAR2, and IL10R2. We show that the large-scale structure of this locus is conserved in human and chicken but not in the pufferfish Fugu rubripes. This establishes that the receptor components of these host defense mechanisms were fixed in an ancestor of the amniotes. The extraordinary diversification of the interferon ligand family during the evolution of birds and mammals has therefore occurred in the context of a fixed receptor structure.  (+info)

Molecular cloning and characterization of the human topoisomerase IIalpha and IIbeta genes: evidence for isoform evolution through gene duplication. (7/1067)

Human DNA topoisomerase II is essential for chromosome segregation and is the target for several clinically important anticancer agents. It is expressed as genetically distinct alpha and beta isoforms encoded by the TOP2alpha and TOP2beta genes that map to chromosomes 17q21-22 and 3p24, respectively. The genes display different patterns of cell cycle- and tissue-specific expression, with the alpha isoform markedly upregulated in proliferating cells. In addition to the fundamental role of TOP2alpha and TOP2beta genes in cell growth and development, altered expression and rearrangement of both genes are implicated in anticancer drug resistance. Here, we report the complete structure of the human topoisomerase IIalpha gene, which consists of 35 exons spanning 27.5 kb. Sequence data for the exon-intron boundaries were determined and examined in the context of topoisomerase IIalpha protein structure comprising three functional domains associated with energy transduction, DNA breakage-reunion activity and nuclear localization. The organization of the 3' half of human TOP2beta, including sequence specifying the C-terminal nuclear localization domain, was also elucidated. Of the 15 introns identified in this 20 kb region of TOP2beta, the first nine and the last intron align in identical positions and display the same phases as introns in TOP2alpha. Though their extreme 3' ends differ, the striking conservation suggests the two genes diverged recently in evolutionary terms consistent with a gene duplication event. Access to TOP2alpha and TOP2beta gene structures should aid studies of mutations and gene rearrangements associated with anticancer drug resistance.  (+info)

Control of fertilization-independent endosperm development by the MEDEA polycomb gene in Arabidopsis. (8/1067)

Higher plant reproduction is unique because two cells are fertilized in the haploid female gametophyte. Egg and sperm nuclei fuse to form the embryo. A second sperm nucleus fuses with the central cell nucleus that replicates to generate the endosperm, a tissue that supports embryo development. To understand mechanisms that initiate reproduction, we isolated a mutation in Arabidopsis, f644, that allows for replication of the central cell and subsequent endosperm development without fertilization. When mutant f644 egg and central cells are fertilized by wild-type sperm, embryo development is inhibited, and endosperm is overproduced. By using a map-based strategy, we cloned and sequenced the F644 gene and showed that it encodes a SET-domain polycomb protein. Subsequently, we found that F644 is identical to MEDEA (MEA), a gene whose maternal-derived allele is required for embryogenesis [Grossniklaus, U., Vielle-Calzada, J.-P., Hoeppner, M. A. & Gagliano, W. B. (1998) Science 280, 446-450]. Together, these results reveal functions for plant polycomb proteins in the suppression of central cell proliferation and endosperm development. We discuss models to explain how polycomb proteins function to suppress endosperm and promote embryo development.  (+info)

Yeah you could use the simple protocl for transformation like one given in manniatis involving cacl2 treated competent cells. I have used this method for transforming 21 kb cosmid. I hope this helps ...
A few weeks ago I described some problems we were having with cosmid libraries. I got several good responses. The one below was the best and might be useful to others. What you have been seeing is actually the gradual loss of the selection pressure, which results from the degradation of the ampicillin in the medium by the beta-lactamase produced by the amp- containing clone. This eventually leads to the loss of the amp- containing cosmid. You might recall the growth of satellite colonies around the genuine transformants when you let the incubation of transformation plates go longer, say two days instead of overnight. After 4-6 weeks the clone has lost the amp-containing cosmid and becomes ampicillin sensitive. That is why you transferred the cells to ampicillin-containing liquid medium and you did not see any growth the next day. The same goes for the lower yield of cosmid DNA after 3-4 weeks, since the colony contains a population of resistant and sensitive cells. My suggestion to you is to ...
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... ,The pWEB::TNC Deletion Cosmid Cloning Kit enables you to prepare, in about two days, a complete and unbiased primary cosmid library of genomic DNA for subsequent use in making nested deletion sublibraries. High efficiency cosmid library construction is accomplished using easy and highly repr,biological,biology supply,biology supplies,biology product
10 mg transfection-grade endofree plasmid or cosmid DNA, HiSpeed Mega, HiSpeed Giga, EF plasmid, HiSpeed Mega and Giga EF Plasmid Kits
The DHFR-G8 cell line was produced by M.C. Hung, et al., in 1986 from the NIH/3T3 mouse fibroblast cell line which was cotransfected with the cNEU-p clone and the pSV2-DHFR plasmid. This cell line was developed from one methotrexate resistant colony by selection in 0.6 uM methotrexate and 10% dialyzed calf serum. These cells contain 50-100 copies of the cNeu-P (normal rat cosmid DNA) and produce approximately 4 x 10(5) molecules of encoded p185 protein per cell.
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cosmid definition: Genetics A hybrid vector that has been spliced with plasmid DNA for cloning huge genes or gene fragments.; a kind of plasmid (often utilized as a cloning vector) constructed because…
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The method used for the isolation of large scale cosmid and plasmid DNA is an unpublished modification (16) of an alkaline lysis procedure (17,18) followed by equilibrium ultracentrifugation in cesium chloride-ethidium bromide gradients (1). Briefly, cells containing the desired plasmid or cosmid are harvested by centrifugation, incubated in a lysozyme buffer, and treated with alkaline detergent. Detergent solubilized proteins and membranes are precipitated with sodium acetate, and the lysate is cleared first by filtration of precipitate through cheesecloth and then by centrifugation. The DNA-containing supernatant is transferred to a new tube, and the plasmid or cosmid DNA is precipitated by the addition of polyethylene glycol and collected by centrifugation. The DNA pellet is resuspended in a buffer containing cesium chloride and ethidium bromide, which is loaded into polyallomer tubes and subjected to ultracentrifugation overnight. The ethidium bromide stained plasmid or cosmid DNA bands, ...
cosmid (16) Duration: 0:14:06 Resolution: 1280x720 Format: wmv Size: 321Mb Download cosmid (16) Download Part 1 From Flyfiles Download Part 2 From
The NucleoBond BAC 100 Kit is designed to purify large DNA fragments such as cosmids, bacteriophage P1 clones, PACs, and BACs, without phenol/chloroform extraction. 1 hour protocol accomodates vectors up to to 300 kb.
Maxiprep NucleoBond Plasmid Kits: Purify plasmid DNA, cosmids, BACs, PACs, and YACs in just a few hours-no phenol/chloroform extraction. Use for transfection, in vitro transcription, and automated sequencing.
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Despite screening various growth media, we failed to detect the production of any 2-hydroxyphenylthiazoline-containing metabolites by S. venezuelae. This is potentially explained by the low levels of sven0516 expression in the bldM mutant (Fig. 3), which is surprising given that sven0517 is likely to be in the same operon, and possibly reflects differential mRNA stability for the two genes. We therefore elected to express the sven0503-sven0517 gene cluster in the engineered host S. coelicolor M1152.23 A clone (SV-2_E03) from an ordered genomic cosmid library of the S. venezuelae chromosome containing a segment extending from sven0496 to sven0518 was PCR-targeted in Escherichia coli with a 5.2 kb SspI fragment from pIJ10702 that contains oriT, and the øC31 integrase gene and phage attachment site (attP). The resulting cosmid, SV-2_E03::SspI, was introduced into S. coelicolor M1152 by conjugation, whereupon it integrated into the chromosomal øC31 attB site. Wild type S. coelicolor M1152 and the ...
TY - JOUR. T1 - Isolation and characterization of a novel gene deleted in digeorge syndrome. AU - Kurahashi, Hiroki. AU - Akagi, Kenzo. AU - Inazawa, Johji. AU - Ohta, Tohru. AU - Niikawa, Norio. AU - Kayatani, Futoshi. AU - Sano, Tetsuya. AU - Okada, Shintaro. AU - Nishisho, Isamu. PY - 1995/4/1. Y1 - 1995/4/1. N2 - The region commonly deleted in DiGeorge syndrome (DGS) has been localized at 22q11.1-q11.2 with the aid of a high resolution banding technique. A 22q11 specific plasmid library was constructed with a microdissection and microcloning method. Dosage analysis proved three of 144 randomly selected microclones to detect hemizygosity in two patients with DGS. Two of the clones were found to contain independent low-copy-number repetitive sequences, all of which were included in the region deleted in the DGS patients. Screening of the cosmid library and subsequent cosmid walking allowed us to obtain two cosmid contigs corresponding to the microclones within the deletion (contig 1 and contig ...
8-base recognition sites will yield pieces 64,000 bases long. Since hundreds of different restriction enzymes have been characterized, DNA can be cut into many different small fragments. Physical Maps Different types of physical maps vary in their degree of resolution. The lowest- resolution physical map is the chromosomal (sometimes called cytogenetic) map, which is based on the distinctive banding patterns observed by light microscopy of stained chromosomes. A cDNA map shows the locations of expressed DNA regions (exons) on the chromosomal map. The more detailed cosmid contig map depicts the order of overlapping DNA fragments spanning the genome. A macrorestriction map describes the order and distance between enzyme cutting (cleavage) sites. The highest- resolution physical map is the complete elucidation of the DNA base- pair sequence of each chromosome in the human genome. Physical maps are described in greater detail below. Low-Resolution Physical Mapping Chromosomal map. In a chromosomal ...
Empirical Bioscience announced the introduction of a new Plasmid Mini-Prep Kit today. Requiring only 1-3mL of bacterial culture, the kit isolates high-quality plasma or cosmid DNA for extraction up to 10 kb in length and yields up to 20 µg per preparation.. In addition to using smaller volumes of cultures, the kit also features an integrated pH indicator within the lysis buffer. This indicator changes color to bright yellow when the lysis buffer reaches the optimal pH level for DNA binding, 7.5. If the pH is greater than 7.5 the lysis buffer will appear orange or violet in color indicating that the pH level is inefficient for DNA adsorption, allowing researchers to make adjustments to the mixture for more efficient plasmid isolation.. Besides the color changing lysis buffer, the kit includes, neutralization buffer, Rnase A, activation buffer, washing buffer, elution buffer, spin columns and collection tubes. These components create a versatile kit that lends itself well for use in ...
The data section can follow or proceed the configuration section. The two sections can also be intermixed. The data section is a tab or whitespace-delimited file which you can export from a spreadsheet application or word processor file (be sure to save as text only!). Here is an example data section:. Cosmid B0511 . 516-619 Cosmid B0511 . 3185-3294 Cosmid B0511 . 10946-11208 Cosmid B0511 . 13126-13511 Cosmid B0511 . 66-208 Cosmid B0511 . 6354-6499 Cosmid B0511 . 13955-14115 EST yk595e6.5 + 3187-3294 EST yk846e07.3 - 11015-11208 EST yk53c10 yk53c10.5 + 18892-19154 yk53c10.3 - 15000-15500,15700-15800 EST yk53c10.5 + 16032-16105 SwissProt PECANEX + 13153-13656 Swedish fish FGENESH Gene 1 - 1-205,518-616,661-735,3187-3365,3436-3846 Transmembrane domain FGENESH Gene 2 - 16626-17396,17451-17597 Kinase and sushi domains. Each line of the file contains five columns. The columns are:. ...
Cosmid clones containing alpha 1-antitrypsin (alpha 1AT) gene sequences were observed to contain alpha 1AT-like sequences approximately 12 kb downstream of the authentic alpha 1AT gene. Restriction mapping suggested the alpha 1AT-like gene lacks promoter sequences. Cosmid clones from one library con …
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Its another Monday and time to introduce another brand new Cosmid girl to you. Her name is Britta and she is from California. She is a 20 year old student that has a serious artsy vibe to her, front what I can tell though, she seems to be a little earthy, if you know what I mean. She is super nice and has such a great smile. She is 57" as well, so a pretty tall drink of water and have a very well proportioned body. I hope that you enjoy out new cutie.. ...
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TY - JOUR. T1 - Detection of submicroscopic chromosomal deletions in aniridia patients using fluorescence in situ hybridization and a panel of cosmids covering the WT1 gene. AU - Kempski, H.. AU - Cowell, J. K.. PY - 1993/1/1. Y1 - 1993/1/1. N2 - A series of cosmids have been isolated from a human chromosome 11-specific cosmid library using the human Wilms tumour predisposition gene cDNA, WT33. Seven overlapping cosmids were isolated which cover the genomic sequence of WT1 and in situ hybridisation shows that they all localise to the p13 region of chromosome 11. Chromosomes from patients with aniridia and Wilms tumour, and a small subband deletion in 11p13, were analysed and no hybridisation signal was seen on the deletion chromosomes. These cosmids, therefore, can be used to analyse chromosomes from patients with sporadic aniridia for submicroscopic deletions. Aniridia patients who show normal hybridisation patterns on both chromosomes need no longer be screened for Wilms tumours.. AB - A ...
The I-2 locus in tomato confers resistance to race 2 of the soil-borne fungus Fusarium oxysporum f sp lycopersici. The selective restriction fragment amplification (AFLP) positional cloning strategy was used to identify I-2 in the tomato genome. A yeast artificial chromosome (YAC) clone covering ~750 kb encompassing the I-2 locus was isolated, and the AFLP technique was used to derive tightly linked AFLP markers from this YAC clone. Genetic complementation analysis in transgenic R1 plants using a set of overlapping cosmids covering the I-2 locus revealed three cosmids giving full resistance to F. o. lycopersici race 2. These cosmids shared a 7-kb DNA fragment containing an open reading frame encoding a protein with similarity to the nucleotide binding site leucine-rich repeat family of resistance genes. At the I-2 locus, we identified six additional homologs that included the recently identified I-2C-1 and I-2C-2 genes. However, cosmids containing the I-2C-1 or I-2C-2 gene could not confer ...
Empirical Bioscience announced the introduction of a new Plasmid Mini-Prep Kit today. Requiring only 1-3mL of bacterial culture, the kit isolates high-quality plasma or cosmid DNA for extraction up to 10 kb in length and yields up to 20 µg per preparation. In addition to using smaller volumes of cultures, the kit also features an integrated pH indicator within the lysis buffer. This indicator changes color to bright yellow when the lysis buffer reaches the optimal pH level for DNA binding, 7.5. If the pH is greater than 7.5 the lysis buffer will appear orange or violet in color indicating that the pH level is inefficient for DNA adsorption, allowing researchers to make adjustments to the mixture for more efficient plasmid isolation. Besides the color changing lysis buffer, the kit includes, neutralization buffer, Rnase A, activation buffer, washing buffer, elution buffer, spin columns and collection tubes. These components create a versatile kit that lends itself well for use in ...
A series of 130 DNA probes, hybridized to the CIC YAC library, are ordered along the abscissa. A subset of 58 of the probes, mapped to chromosome 2 by RFLP analysis, serve as contact points between the genetic and physical maps. The positions of markers mapped to the recombinant inbred lines (7) are indicated in cM above the marker names. When a set of probes hybridized to the same YACs, their order is listed by their position in the genetic map (7). If no genetic or auxilliary data (e.g. cosmid contig (4)) was available, the exact probe order is indeterminate and is arbitrarily assigned.. A line drawn above a marker indicates a difference in marker order between the physical and genetic maps. Markers designated "RI" have been mapped by us to the recombinant inbred populations and are in correct relative position (with one exception, 15414, marked "RI-?") to the markers on either side (no number is indicated as adding markers and using a different mapping program generates different absolute ...
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Brian Robertson wrote: , , The amount of bacterial genome data available as sequenced cosmids of , 30-40 kb is increasing rapidly. Our problem is that we need to keep track , of newly discovered genes as they appear, so they can be incorporated into , our research program as appropriate. For this we need to create lists of , probable genes identified in the annotations for each cosmid. This can , then be circulated to laboratory workers. , , An example of this kind of annotation is shown below. We would like to , extract the /note field, which contains the probable function of the , gene, and create a list of these for each cosmid. , , FT CDS_pept complement(3043..4155) , FT /note=MTCY190.03c, probable anthranilate , FT phosphoribosyltransferase, trpD, len: 370, similar to eg , FT SW:TRPD_LACCA P17170, (43.2% identity in 308 aa overlap), , FT initiation codon uncertain, gtg at 4086 favoured by , FT homology but this has no clear ribosome binding site , , Does anyone know of a way of ...
The kits utilize a novel strategy of cloning end-repaired, randomly sheared DNA instead of the conventional approach of cloning fragments generated by partial restriction endonuclease digestion. First, genomic DNA is sheared by passing it through a syringe needle. The sheared DNA is end-repaired to generate 5-phosphorylated blunt ends and size-selected using a low melting point agarose gel. The size-selected DNA is then ligated into the supplied linearized and dephosphorylated pWEB-TNCTM or pWEBTM Cosmid Vector, packaged using ultra-high efficiency MaxPlaxTM Lambda Packaging Extracts (,109 pfu/μg for phage lambda) and plated on phage T1-resistant EPI100TM-T1R E. coli plating cells, all included in the kit. The result is a complete and unbiased primary cosmid library ...
The kits utilize a novel strategy of cloning end-repaired, randomly sheared DNA instead of the conventional approach of cloning fragments generated by partial restriction endonuclease digestion. First, genomic DNA is sheared by passing it through a syringe needle. The sheared DNA is end-repaired to generate 5-phosphorylated blunt ends and size-selected using a low melting point agarose gel. The size-selected DNA is then ligated into the supplied linearized and dephosphorylated pWEB-TNCTM or pWEBTM Cosmid Vector, packaged using ultra-high efficiency MaxPlaxTM Lambda Packaging Extracts (,109 pfu/μg for phage lambda) and plated on phage T1-resistant EPI100TM-T1R E. coli plating cells, all included in the kit. The result is a complete and unbiased primary cosmid library ...
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From a cosmid gene bank of Bacillus cereus GP4 in Escherichia coli we isolated clones which, after several days of incubation, formed hemolysis zones on erythrocyte agar plates. These clones contained recombinant cosmids with B. cereus DNA insertions of varying lengths which shared some common restriction fragments. The smallest insertionwas recloned as aPstl fragment into pJKK3-1, a shuttle vector which repücates in Bacillus subtilis and E. coli. When this recombinant plasmid (pJKK3-1 hly-1) was transformed into E. coli, it caused hemolysis on erythrocyte agar plates, but in liquid assays no extemal or intemal hemolytic activity could be detected with the E. coli transformants. B. subtilis carrying the same plasmid exhibited hemolytic activity at Ievels comparable to those ofthe B. cereus donor strain. The hemolysin produced in B. subtilis seemed to be indistinguishable from cereolysin in its sensitivity to cholesterol, activation by dithiothreitol, and inactivation by antibodies raised ...
Fosmids are similar to cosmids but are based on the bacterial F-plasmid. The cloning vector is limited, as a host (usually E. coli) can only contain one fosmid molecule. Fosmids can hold DNA inserts of up to 40 kb in size; often the source of the insert is random genomic DNA. A fosmid library is prepared by extracting the genomic DNA from the target organism and cloning it into the fosmid vector. The ligation mix is then packaged into phage particles and the DNA is transfected into the bacterial host. Bacterial clones propagate the fosmid library. The low copy number offers higher stability than vectors with relatively higher copy numbers, including cosmids. Fosmids may be useful for constructing stable libraries from complex genomes. Fosmids have high structural stability and have been found to maintain human DNA effectively even after 100 generations of bacterial growth. Fosmid clones were used to help assess the accuracy of the Public Human Genome Sequence. The fertility plasmid or F-plasmid ...
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A long-range physical map of the carcinoembryonic antigen (CEA) gene family cluster, which is located on the long arm of chromosome 19, has been constructed. This was achieved by hybridization analysis of large DNA fragments separated by pulse-field gel electrophoresis and of DNA from human/rodent somatic cell hybrids, as well as the assembly of ordered sets of cosmids for this gene region into contigs. The different approaches yielded very similar results and indicate that the entire gene family is contained within a region located at position 19q13.1-q13.2 between the CYP2A and the D19S15/D19S8 markers. The physical linkage of nine genes belonging to the CEA subgroup and their location with respect to the pregnancy-specific glycoprotein (PSG) subgroup genes have been determined, and the latter are located closer to the telomere. From large groups of ordered cosmid clones, the identity of all known CEA subgroup genes has been confirmed either by hybridization using gene-specific probes or by ...
In enzymatic sequencing by primer walking, a short, known DNA sequence is used with the enzyme DNA polymerase to "prime" or trigger the synthesis of a new DNA strand complementary to a template of unknown sequence. The sequence of about 500 bases on the new chain can be determined with standard gel electrophoresis methods, and a short stretch at the end of the newly determined sequence is then used to design another primer to extend or "walk" down the next 500 bases. Successive primers selected in this way are used to determine an entire unknown DNA sequence.. The authors believe primer walking is the best strategy for sequencing cosmids (clones containing about 40,000 bp of DNA). The preparation of only one DNA sample is needed to sequence an entire cosmid, without the need for subcloning or multiple template preparation. Primer walking offers low redundancy and high accuracy, and sequence assembly is straightforward. Sequencing of problem areas is facilitated by the freedom to select almost ...
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The most variable human minisatellites show extreme germline instability dominated by complex intra-allelic rearrangements plus a lower frequency of inter-allelic transfers of repeat units. In contrast, little is known about somatic instability at such loci. We have therefore used single-molecule PCR to analyze mutation at minisatellite CEB1 (D2S90) in human blood DNA. Somatic mutants were rare and involved only relatively simple intra-allelic events, with no bias toward expansions, in sharp contrast to the complex gain-biased rearrangements seen in sperm. Somatic and germline mutation processes were further analyzed in mice transgenic for a cosmid insert containing CEB1. Mutant molecules in transgenic sperm and blood were detected but only at the low frequencies seen in human blood and arose mainly by simple duplications and deletions as seen for somatic mutations in human. These data suggest distinct pathways for germline and somatic CEB1 mutations with germline instability involving ...
A 2 year old female presenting with bilateral sporadic aniridia was found to have an apparently balanced reciprocal translocation with a chromosome 11 breakpoint within band p13. Fluorescence in situ hybridisation (FISH) studies with distal 11p13 specific cosmids showed that the chromosome 11 breakpoint lay between the aniridia (PAX6) locus and a region approximately 100 kb distal to PAX6 defined by the cosmid FO2121. Although this patient did not have a detectable deletion within PAX6, her aniridia may have resulted from a disruption of the distal chromatin domain containing either enhancers or regulators for PAX6. This case may therefore be another example of aniridia caused by a position effect as recently described in two familial aniridia patients in which the phenotype cosegregated with chromosome abnormalities with 11p13 breakpoints.. ...
We constructed cosmid and fosmid libraries of Sorangium cellulosum So0157-2 genome. Southern blotting hybridization with epothilone gene probes revealed clones containing the epothilone genes. The harbored epothilone genes fragments in the positive clones were determined by the end sequencing. We selected Cosmid10 and Fosmid3B11 for our construction. After completely sequencing, Cosmid10 contained a fragment of complete epoA to epoC and part of epoD, while Fosmid3B11 contained part of epoC till the downstream sequence of epoF.. To stitch the two epothilone-genes fragments in Cosmid10 and Fosmid3B11, transitional recombinant vectors p15A- recT and p15A-recM were constructed as follows. First, the 2.0-kb BglII-XbaI epoup fragment (upstream of epoA), 2.2-kbXbaI-NdeI epoCov fragment (overlapping the epoC sequence), and the 2.2-kb NotI-SpeI epodown fragment (downstream of epoF) were amplified from the genomic DNA of S. cellulosum So0157-2 using the primer pairs of UP, C-LAP and DOWN, respectively. ...
The t-haplotype is a chromosomal region in Mus musculus characterized by meiotic drive such that heterozygous males transmit t-bearing chromosomes to roughly 90% of their offspring. Most naturally occurring t-haplotypes express a recessive embryonic lethality, preventing fixation of the t-haplotype. Surprisingly, the t-haplotype occurs in nature as a persistent, low-frequency polymorphism. Early modeling studies led LEWONTIN to hypothesize that this low level polymorphism results from a balance between genetic drift in small demes and interdemic migration. Here, we show that while combination of deme size and migration rate that predict natural t-haplotype frequencies exist, the range of such values is too narrow to be biologically plausible, suggesting that small deme size and interdemic migration alone do not explain the observed t-haplotype frequencies. In response, we tested other factors that might explain the observed t-polymorphism. Two led to biologically plausible models: substantially reduced
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Digitoxosyltetracenomycin C and Glucosyltetracenomycin C, Two Novel Elloramycin Analogues Obtained by Exploring the Sugar Donor Substrate Specificity of Glycosyltransferase ElmGT
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The third component of complement (C3) is an important mediator of inflammation. Murine and human genomic cosmid clones were isolated, characterized and sequenced 5′ to the complement C3 gene transcriptional initiation sites to determine cis elements that participate in constitutive and regulated C3 gene expression. The murine and human 5′ flanking regions are 51% identical overall, with positions -36 to -1 and -146 to -68 showing 80% identity. Four TATA boxes were identified upstream of the murine transcriptional initiation site, but deletion and transfection analysis using reporter gene constructs in HepG2 cells indicated that only the TATA element at position -30, together with sequences -395 to -111, are essential for constitutive expression of murine C3 in hepatocytes. Deletion analysis also suggested that sequences between -1457 and -800 contain regulatory elements that are involved in suppressing basal expression. Sequences between -90 to -41 confer both enhancer activity and ...
Birkenbihl, R.P.; Vielmetter, W., 1989: Cosmid derived map of escherichia coli strain bhb2600 in comparison to the map of strain w3110
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1. Rabbits dosed orally with β-ionone excreted in the urine unchanged β-ionone, 3-oxo-β-ionone, 3-oxo-β-ionol, dihydro-3-oxo-β-ionol and 3-hydroxy-β-ionol. 2. Excretion products were isolated as 2,4-dinitrophenylhydrazone derivatives (β-ionone, 3-oxo-β-ionone, 3-oxo-β-ionol and dihydro-3-oxo-β-ionol) and as p-nitrobenzoate derivatives (3-oxo-β-ionol, dihydro-3-oxo-β-ionol and 3-hydroxy-β-ionol), which were characterized and identified by comparison with the synthetic authentic compounds. 3. The glucuronides of 3-oxo-β-ionol and dihydro-3-oxo-β-ionol were also detected in the urine. The latter compound was isolated as free glucuronide, sodium salt and 2,4-dinitrophenylhydrazone.. ...
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Plasmid Cosmid Fosmid Katoh, Motonobu; Ayabe, Fumiaki; Norikane, Satoko; Okada, Teruaki; Masumoto, Hiroshi; Horike, Shin-iche; ...
Once inside the host, the cosmids circularize with the aid of the host's DNA ligase and then function as plasmids. Cosmids are ... This sequence allows the cosmid to be packaged into bacteriophage λ particles. These particles- containing a linearized cosmid ... Cosmid vectors are plasmids that contain a small region of bacteriophage λ DNA called the cos sequence. ...
"Physical mapping of the mitochondrial genome of Arabidopsis thaliana by cosmid and YAC clones". Plant Journal. 6 (3): 447-455. ...
Alu repeats and polymorphisms in cosmids sequenced from chromosome 4p16.3". Nature Genetics. 1 (5): 348-53. doi:10.1038/ng0892- ...
... s are similar to cosmids but are based on the bacterial F-plasmid. The cloning vector is limited, as a host (usually E. ... The major advantage of Fosmids over other cosmid systems lies in its capability of stably propagating human DNA fragments. ... 1992). "Stable propagation of cosmid-sized human DNA inserts in an F-factor based vector". Nucleic Acids Res. 20 (5): 1083-1085 ... The low copy number offers higher stability than vectors with relatively higher copy numbers, including cosmids. Fosmids may be ...
The four major types of vectors are plasmids, viral vectors, cosmids, and artificial chromosomes. Of these, the most commonly ... cosmid, Lambda phages). A vector containing foreign DNA is termed recombinant DNA. ...
... indicating it is on the cosmid 'F38H4', and there are at least 6 other genes on that cosmid. If a gene produces a protein that ... For example, in 2005 a 39 kb cosmid had to be inverted. Other improvements have come from comparing genomic DNA to cDNA ... These species therefore do not have sequence names for CDSs and gene transcripts that are based on cosmid names. Instead they ... which is derived from the cosmid, fosmid or YAC clone on which they reside, for instance F38H4.7, ...
It is encoded by the P87050 2247 bp ORF on the cosmid 57A10. The protein is 775 amino acids in length. Cdr2 is a member of the ...
1982). "Isolation of human oncogene sequences (v-fes homolog) from a cosmid library". Science. 216 (4550): 1136-8. doi:10.1126/ ...
1990). "High-resolution mapping of human chromosome 11 by in situ hybridization with cosmid clones". Science. 247 (4938): 64-9 ...
1994). "Isolation of cosmid and cDNA clones in the region surrounding the BTK gene at Xq21.3-q22". Genomics. 21 (3): 517-24. ...
... a 1 Mbp cosmid and PAC contig". Genes, Chromosomes & Cancer. 31 (3): 228-39. doi:10.1002/gcc.1139. PMID 11391793. Ruppert JM, ...
1999). "A 500-kb sequence-ready cosmid contig and transcript map of the MEN1 region on 11q13". Genomics. 55 (1): 49-56. doi: ...
Cosmid/BAC/YAC end sequences use Cosmid/Bacterial artificial chromosome/Yeast artificial chromosome to sequence the genome from ... Cosmid/BAC/YAC can also be used to get bigger clone of DNA fragment than vectors like plasmid and phagemid. A larger insert is ... BAC can't do that, but BACs can reliably represent human DNA much better than YAC or cosmid. Exon trapped sequence is used to ... DDBJ-GSS MEGA- and GIGA preps of cosmid-, BAC-, PAC, YAC-, and P1-DNA with JETSTAR 2.0 WSSP-04 Chapter 2 - Vectors Yeast ...
1994). "Isolation of cosmid and cDNA clones in the region surrounding the BTK gene at Xq21.3-q22". Genomics. 21 (3): 517-24. ...
The sequence was obtained by a combined approach, employing automated DNA sequence analysis of selected cosmids and whole- ...
... cytogenetic band location of 540 cosmids and 70 genes or DNA markers". Genomics. 15 (1): 133-45. doi:10.1006/geno.1993.1021. ...
Mar 1993). "Fluorescence in situ hybridization mapping of human chromosome 19: cytogenetic band location of 540 cosmids and 70 ...
Cosmid End-sequence profiling Fosmid Human artificial chromosome Yeast artificial chromosome O'Connor M, Peifer M, Bender W ( ...
1996). "A set of ordered cosmids and a detailed genetic and physical map for the 8 Mb Streptomyces coelicolor A3(2) chromosome ...
1993). "Fluorescence in situ hybridization mapping of human chromosome 19: cytogenetic band location of 540 cosmids and 70 ...
A set of ordered cosmids and a detailed genetic and physical map for the 8 Mb Streptomyces coelicolor A3(2) chromosome. Mol ...
... five cosmids should be used for microdeletion detection and low number of protein truncating mutations". Journal of Medical ...
For instance, phage TM4 was used to construct shuttle phasmids that replicate as large cosmids in Escherichia coli and as ...
Phil Green for the assembly of cosmids in large-scale cosmid shotgun sequencing within the Human Genome Project. Phrap has been ... Typically, Phrap had no problems differentiating between the different Alu copies in a cosmid, and to correctly assemble the ... cosmids (or, later, BACs). The logic is simple: a base call with a high probability of being correct should never be aligned ...
... cells and we can make good cosmid DNA from them. However, after 3-4 weeks , the yield of cosmid DNA from cells derived from the ... It is NO NEED to pick up a single colony for your cosmid preparation each time. After you have checked and know that the cosmid ... Has anyone else had this problem? Are we leaving something out of the , media? Are the cosmid-containing cells less viable? , , ... working with cosmids. Barry Bowman bowman at biology.ucsc.edu Tue Jun 20 19:26:42 EST 1995 *Previous message: only test ...
... Dave Fruhling Fruhlid at war.wyeth.com Tue May 9 15:08:03 EST 2000 *Previous message: Sequencing from ...
Medical definition of cosmid: a plasmid into which a short nucleotide sequence of a bacteriophage has been inserted to create a ... Resources for cosmid. Time Traveler: Explore other words from the year cosmid first appeared Time Traveler! Explore the year a ... Comments on cosmid. What made you want to look up cosmid? Please tell us where you read or heard it (including the quote, if ... Post the Definition of cosmid to Facebook Share the Definition of cosmid on Twitter ...
... Denise Melanson dmmelans at unity.ncsu.edu Sat Dec 21 21:25:12 EST 1996 *Previous message: ... I am looking for a cosmid library of A. tumefaciens genome. I dont need the Ti plasmids, I actually want the genomic DNA from ...
A cosmid is a type of hybrid plasmid that contains a Lambda phage cos sequence. Cosmids (cos sites + plasmid = cosmids) DNA ... Depending on the particular aim of the experiment broad host range cosmids, shuttle cosmids or mammalian cosmids (linked to ... Cosmids can be used to build genomic libraries. They were first described by Collins and Hohn in 1978. Cosmids can contain 37 ... Selection against wild type cosmid DNA is simply done via size exclusion. Cosmids, however, always form colonies and not ...
If I want to pick a good cosmid to inject for mutant rescue, how can I tell with acedb which cosmid will have the ORF lying ... I am interested in a predicted gene that seems to be at one end of a sequenced cosmid, perhaps even extending from one cosmid ... Interpreting cosmid overlaps with acedb. Curtis Loer cloer at acusd.edu Mon Aug 25 16:17:22 EST 1997 *Previous message: Cell ... could someone help me with interpreting the fingerprinting info to pick a few good cosmids? Thanks. Curtis M. Loer Dept. of ...
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Cosmid shuttle vectors for cloning and analysis of Streptomyces DNA.. Rao RN, Richardson MA, Kuhstoss S. ...
... "sequence overlap with cosmid g8261" misc_feature 7505..7506 /note="site of apparent internal deletion in cosmid" BASE COUNT ... REFERENCE 1 (bases 1 to 11862) AUTHORS Goodman,H.M. and Gallant,P. TITLE Sequence of cosmid g17311 from Arabidopsis thaliana ... LOCUS ATU53502 11862 bp DNA PLN 05-MAY-1996 DEFINITION Arabidopsis thaliana chromosome I cosmid g17311 DNA. ACCESSION U53502 ...
LOCUS ATU53501 37570 bp DNA PLN 06-MAY-1996 DEFINITION Arabidopsis thaliana chromosome I cosmid g8261 DNA (cytosine-5-) ...
TNC Deletion Cosmid Cloning Kit enables you to prepare, in about two days, a complete and unbiased primary cosmid library of ... High efficiency cosmid library construction is accomplished using easy and highly repr,biological,biology supply,biology ... TNC Deletion Cosmid Transposition Kit from EPICENTRE Biotechnologies,The pWEB:: ... The pWEB::TNC Deletion Cosmid Cloning Kit enables you to prepare, in about two days, a complete and unbiased primary cosmid ...
One cosmid in set A contains the oriL locus in a nonoverlapping region and lacks ori+L forms. This set generated only ori-L ... Two cosmids common to sets B and C ostensibly contain one of the origins of viral DNA replication (oriL) in a region of overlap ... A cosmid-based system for constructing mutants of herpes simplex virus type 1.. Cunningham C1, Davison AJ. ... Cosmids containing large fragments of herpes simplex virus type 1 DNA were prepared using a vector that allows intact inserts ...
Complementation of a Chlamydomonas reinhardtii mutant using a genomic cosmid library.. Purton S1, Rochaix JD. ... of Chlamydomonas reinhardtii by complementation using total DNA from a genomic cosmid library. Using the glass-bead ...
250 people for cosmid: High School, Busty, Babes, Galleries, TX, CA, Videos, Pics Galleries, Profile, University, free people ... 05.02.2012 Jeannine Hansen Cosmid Jeannine Hansen Cosmid Jeannine Hansen Cosmid Jeannine Hansen Cosmid Jeannine Hansen Cosmid ... More of Holly Riley and other busty girls next door at Cosmid · Holly Riley Cosmid 1 Holly Riley Cosmid 2 Holly Riley Cosmid 3 ... Christine carollo cosmidhristine carollo cosmid: Christine carollo cosmidhristine carollo cosmid. Relaxinol ingredients. You ...
Cosmids were provided by the European Drosophila Genome Project (EDGP).. EDGP cosmids are available from Geneservice in the UK ... Cosmids are available either singly or on high-density filters.. The members of the European Drosophila Genome mapping ... Contact for cosmids:. Dr. I. Siden-Kiamos, Institute of Molecular Biology and Biotechnology, FORTH, Heraklion, Crete. Telephone ...
A 1.7-megabase sequence-ready cosmid contig covering the TSC1 candidate region in 9q34.. Hornigold N1, van Slegtenhorst M, ...
When plasmids or cosmids are larger than 10 kb, pre-heat Buffer EB (or water) to 70°C prior to eluting DNA from the QIAprep ... All QIAprep Miniprep Kits can be used for preparation of low-copy number plasmids and cosmids up to 50 kb. Below are ... Can I use QIAprep Miniprep kits for low-copy plasmids and cosmids? ...
Final sequence length of the cosmid is 37570 bp.. Assembly of the cosmid sequence g17311 revealed that the cosmid had suffered ... These overlapping cosmids are part of a series of approximately 17,000 cosmids which we have fingerprinted (1, 2) and some of ... Thus in a single plate there are 96 x N (N is the number of vectors used, which was 7 in our analysis of cosmids g8261 and ... Cosmid g8261 was sequenced, but the ,7 fold sampling redundancy achieved did not produce complete coverage. The small gaps or ...
One colony: One library cosmid DNA miniprep. Because of the instability of the Olson cosmids, we have divised a miniprep ... Both the cosmid DNA and the E. coli DNAare denatured. However, only the cosmid DNA is supercoiled. Its two strands cannot ... The amount of cosmid DNA prepared is modest, but is more than enough to make an M13 library. While most genomists focus on the ... If you dont select the right E. coli colony, you cannot prepare the right cosmid DNA. 1-Late in your working day streak ...
Misc.Comments : Suicide vector permitting gene replacement and mobilization into a wide range of Gram negative bacteria. Allows positive selection for integration. Replicates only in enterobacteria, and so functions as a suicide vector in other Gram negative hosts. When induced by media containing 5% sucrose, sacB is lethal in a wide range of Gram negative bacteria, and thus permits selection for loss of the vector. A single BglII site within the gentamicin resistance gene (gtmR) allows insertion of other resistance genes (e.g., as a BamHI cassette). The order of the major features of the plasmid is: gtmR - EcoRI/MCS/HindIII - P15A - cos - traJ - oriT - sacB. [1] Deposited by: M.F.Hynes Grwoth media: LB plus gentamicin (ATCC medium number 1885) 37C ...
The pWEB™ Cosmid Cloning Kit facilitates rapid and efficient construction of cosmid libraries using the pWEB Cosmid Vector (Fig ... pWEB™ Cosmid Cloning Kit. Notice: The pWEB Cosmid Cloning Kit has been discontinued.. As alternative, please consider the ... Figure 2 (click to enlarge). Outline of the cosmid cloning procedure using the pWEB-TNC™ and the pWEB™ Cosmid Cloning Kits.. ... The result is a complete and unbiased primary cosmid library.. Epicentre also offers kits for fosmid cloning and library ...
pLAFR1 cosmids were mobilized from R. meliloti prototrophic colonies into E. coli and then reintroduced into R. meliloti ... Construction of a broad host range cosmid cloning vector and its use in the genetic analysis of Rhizobium mutants.. Friedman AM ... We have constructed a cosmid derivative of the low copy-number broad host-range cloning vector pRK290 (Ditta et al., 1980) by ...
how to get cosmid DNA back to E.coli? - (Aug/07/2006 ). Hi,. I need to know if it is possible to transform E.coli with cosmid ... I need to know if it is possible to transform E.coli with cosmid vector DNA (approx 30 kb)?. If anyone has a protocol or a ... I need to know if it is possible to transform E.coli with cosmid vector DNA (approx 30 kb)?. If anyone has a protocol or a ... I have used this method for transforming 21 kb cosmid. I hope this helps.. uhsna. quote name=puma date=Aug 7 2006, 12:27 PM ...
Epstein-Barr virus recombinants from overlapping cosmid fragments.. B Tomkinson, E Robertson, R Yalamanchili, R Longnecker, E ... Epstein-Barr virus recombinants from overlapping cosmid fragments.. B Tomkinson, E Robertson, R Yalamanchili, R Longnecker, E ... Epstein-Barr virus recombinants from overlapping cosmid fragments.. B Tomkinson, E Robertson, R Yalamanchili, R Longnecker, E ... Epstein-Barr virus recombinants from overlapping cosmid fragments. Message Subject (Your Name) has forwarded a page to you from ...
cosmid definition: Genetics A hybrid vector that has been spliced with plasmid DNA for cloning huge genes or gene fragments.; a ... Definition for "cosmid"*Genetics A hybrid vector that has been spliced… ... How would you define cosmid?. All the definitions on AZdictionary were written by people just like you. Nows your chance to ... Sentence for "cosmid"*E. coli and 25 µg/ml for… ... Medical Dictionary for "cosmid"*DNA from a bacterial ...
  • Cosmids are predominantly plasmids with a bacterial oriV, an antibiotic selection marker and a cloning site, but they carry one, or more recently two, cos sites derived from bacteriophage lambda. (wikipedia.org)
  • Can I use QIAprep Miniprep kits for low-copy plasmids and cosmids? (qiagen.com)
  • All QIAprep Miniprep Kits can be used for preparation of low-copy number plasmids and cosmids up to 50 kb. (qiagen.com)
  • Thus cosmids are essentially plasmids which have had a small piece of bacteriophage lambda DNA including the cos site (that is, containing the single stranded cohesive/sticky endings of λ phage which are complimentary to each other) cloned into them (Lodge, Lund & Minchin, 2007). (bartleby.com)
  • The development at Oncor of a Her-2/neu cosmid clone suitable for fluorescence in situ hybridization is an important new development. (freethesaurus.com)
  • Kempski, H & Cowell, JK 1993, ' Detection of submicroscopic chromosomal deletions in aniridia patients using fluorescence in situ hybridization and a panel of cosmids covering the WT1 gene ', International journal of oncology , vol. 3, no. 5, pp. 937-940. (elsevier.com)
  • When the clone bank was mated (en masse) from Escherichia coli to various R. meliloti auxotrophic mutants, tetracycline-resistant (Tcr) transconjugants were obtained at frequencies ranging from 0.1 to 0.8, and among these, prototrophic colonies were obtained at frequencies ranging from 0.001 to 0.007. (nih.gov)
  • Two pathogenicity islands in uropathogenic Escherichia coli J96: cosmid cloning and sample sequencing. (asm.org)
  • Created with DNA from a bacteriophage (a virus that infectsbacteria) combined with genetic material called a cosmid from the common bacterium Escherichia coli, the shuttle phasmid can infect mycobacteria and grow in E. (freethesaurus.com)
  • For instance, phage TM4 was used to construct shuttle phasmids that replicate as large cosmids in Escherichia coli and as phages in mycobacteria. (wikipedia.org)
  • Epstein-Barr virus recombinants from overlapping cosmid fragments. (asm.org)
  • These recombinants probably result from recombination among the transfected cosmid-cloned EBV DNA fragments. (asm.org)
  • Of the recombinants which had incorporated the marker-rescuing cosmid DNA fragment and the fragment encoding the type 1 EBNA 3A gene, most had incorporated markers from at least two other transfected cosmid DNA fragments, indicating a propensity for multiple homologous recombinations. (asm.org)
  • The major advantage of Fosmids over other cosmid systems lies in its capability of stably propagating human DNA fragments. (wikipedia.org)
  • The de novo generated circles had exact multiples of tandem copies of 2-kb fragments from cosmid templates and that the tandem organization was a prerequisite to circle formation. (wikipedia.org)
  • Viral mutants with lesions in either or both of genes UL2 and UL44, which are not essential for growth in cell culture, were constructed using cosmids containing specifically introduced frameshift mutations. (nih.gov)
  • EBV recombinants which have the type 1 EBNA LP and 2 genes from the transfected EcoRI-A cosmid DNA were selectively and clonally recovered by exploiting the unique ability of the recombinants to transform primary B lymphocytes into lymphoblastoid cell lines. (asm.org)
  • We provide evidence that they are induced by introducing the cosmid St4A10∆lsp, as part of ReDirect PCR mutagenesis protocol, which transiently duplicates a number of important cell division genes. (northumbria.ac.uk)
  • These cosmids shared a 7-kb DNA fragment containing an open reading frame encoding a protein with similarity to the nucleotide binding site leucine-rich repeat family of resistance genes. (plantcell.org)
  • It is a cosmid, and contains two functioning (wild-type) closely linked genes: a gene of interest and a mosaic marker. (wikipedia.org)
  • cells and we can make good cosmid DNA from them. (bio.net)
  • If I want to pick a good cosmid to inject for mutant rescue, how can I tell with acedb which cosmid will have the ORF lying more-or-less in the center instead of at the extreme of one end? (bio.net)
  • Complementation of a Chlamydomonas reinhardtii mutant using a genomic cosmid library. (nih.gov)
  • We report the rescue of an arginine-requiring mutant (arg7-8) of Chlamydomonas reinhardtii by complementation using total DNA from a genomic cosmid library. (nih.gov)
  • Because of the instability of the Olson cosmids, we have divised a miniprep procedure that minimizes E. coli growth, thus allowing the least possible time for cosmid DNA deletions/rearrangements to arise and be selected. (stanford.edu)
  • If you don't select the 'right' E. coli colony, you cannot prepare the 'right' cosmid DNA. (stanford.edu)
  • If you see all large colonies, proceed with caution, as you may have E. coli containing a cosmid with deletions. (stanford.edu)
  • First thing in the morning, harvest the E. coli and prepare cosmid DNA. (stanford.edu)
  • Both the cosmid DNA and the E. coli DNAare denatured. (stanford.edu)
  • When the solution is brought to neutral pH, the cosmid DNA renatures spontaneously, while the E. coli DNA does not renature under these conditions. (stanford.edu)
  • pLAFR1 cosmids were mobilized from R. meliloti prototrophic colonies into E. coli and then reintroduced into R. meliloti auxotrophs. (nih.gov)
  • how to get cosmid DNA back to E.coli? (protocol-online.org)
  • One cosmid in set A contains the oriL locus in a nonoverlapping region and lacks ori+L forms. (nih.gov)
  • Genetic complementation analysis in transgenic R 1 plants using a set of overlapping cosmids covering the I2 locus revealed three cosmids giving full resistance to F. o. lycopersici race 2. (plantcell.org)
  • These results indicate that a cosmid-based system will facilitate isolation of large numbers of defined viral mutants. (nih.gov)
  • These overlapping cosmids are part of a series of approximately 17,000 cosmids which we have fingerprinted ( 1, 2 ) and some of which have been mapped as RFLPs ( 3 ) onto the five chromosomes of Arabidopsis. (harvard.edu)
  • These cosmids, therefore, can be used to analyse chromosomes from patients with sporadic aniridia for submicroscopic deletions. (elsevier.com)
  • P3HR-1 cells harbor a type 2 EBV which is unable to transform primary B lymphocytes because of a deletion of DNA encoding EBNA LP and EBNA 2, but the P3HR-1 EBV can provide replication functions in trans and can recombine with the transfected cosmids. (asm.org)
  • BAC can't do that, but BACs can reliably represent human DNA much better than YAC or cosmid. (wikipedia.org)
  • Typically, Phrap had no problems differentiating between the different Alu copies in a cosmid, and to correctly assemble the cosmids (or, later, BACs). (wikipedia.org)
  • Depending on the particular aim of the experiment broad host range cosmids, shuttle cosmids or 'mammalian' cosmids (linked to SV40 oriV and mammalian selection markers) are available. (wikipedia.org)
  • This causes linearization of the circular cosmid with two "cohesive" or "sticky ends" of 12bp. (wikipedia.org)
  • Cosmid - Als Cosmide bezeichnet man Plasmide, die sogenannte cos sites (cohesive site) enthalten. (enacademic.com)
  • The cosQ site of next cosmid (as rolling circle replication often results in linear concatemers) is held by the terminase after the previous cosmid has been packaged, to prevent degradation by cellular DNases. (wikipedia.org)
  • The chromosomal localization of the cloned exons was inferred from the localization of the parent cosmids. (deepdyve.com)
  • The one recombinant virus examined in detail by Southern blot analysis has all the polymorphisms characteristic of the transfected type 1 cosmid DNA and none characteristic of the type 2 P3HR-1 EBV DNA. (asm.org)
  • When either of these cosmids was replaced by a derivative comprising only ori-L forms, ori+L and ori-L progeny were obtained, and only ori-L progeny were produced when both were replaced. (nih.gov)
  • Two cosmids common to sets B and C ostensibly contain one of the origins of viral DNA replication (oriL) in a region of overlap between inserts, but both actually consist of a minority of apparently intact (ori+L) forms and a majority of deleted (ori-L) forms. (nih.gov)
  • In his post-doctoral research he constructed the first reliable method for cloning human DNA cosmids - at the time an extremely difficult but rewarding accomplishment that has since been adopted by many other laboratories. (wikipedia.org)
  • The cosmid, which is not packed correctly during mitosis, is occasionally present in only one daughter cell following cell division. (wikipedia.org)
  • The same goes for the lower yield of cosmid DNA after 3-4 weeks, since the colony contains a population of resistant and sensitive cells. (bio.net)
  • It is NO NEED to pick up a single colony for your cosmid preparation each time. (bio.net)