genomic DNA

About: genomic DNA is a(n) research topic. Over the lifetime, 15046 publication(s) have been published within this topic receiving 663636 citation(s). The topic is also known as: genomic deoxyribonucleic acid & gDNA.

PCR protocols : A guide to methods and applications

Abstract: Basic Methodology: M.A. Innis and D.H. Gelfand, Optimization of PCRs. R.K. Saiki, Amplification of Genomic DNA. E.S. Kawasaki, Amplification of RNA. M.A. Frohman, RACE: Rapid Amplification of cDNA Ends. T. Compton, Degenerate Primers for DNA Amplification. C.C. Lee and C.T. Caskey, cDNA Cloning Using Degenerate Primers. M.A. Innis, PCR with 7-Deaza-2~b7-Deoxyguanosine Triphosphate. G. Gilliland, S. Perrin, and H.F. Bunn, Competitive PCR for Quantitation of mRNA. A.M. Wang and D.F. Mark, Quantitative PCR. P.C. McCabe, Production of Single-Stranded DNA by Asymmetric PCR. S.J. Scharf, Cloning with PCR. U. Landegren, R. Kaiser, and L. Hood, Oligonucleotide Ligation Assay. C. Levenson and C.-A. Chang, Nonisotopically Labeled Probes and Primers. Y-M.D. Lo, W.Z. Mehal, and K.A. Fleming, Incorporation of Biotinylated dUTP. R. Helmuth, Nonisotopic Detection of PCR Products. D.H. Gelfand and T.J. White, Thermostable DNA Polymerases. S. Kwok, Procedure to Minimuze PCR-Product Carry-Over. E.S. Kawasaki, Sample Preparation from Blood, Cells, and Other Fluids. D.K. Wright and M.M. Manos, Sample Preparation from Paraffin-Embedded Tissues. S. P~ada~adabo, Amplifying Ancient DNA. Research Applications. M.J. Holland and M.A. Innis, In Vitro Transcription of PCR Templates. R. Higuchi, Recombinant PCR. B. Krummel, DNase I Footprinting. M.A.D. Brow, Sequencing with Taq DNA Polymerase. S.S. Sommer, G. Sarkar, D.D. Koeberl, C.D.K. Bottema, J.-M. Buerstedde, D.B. Schowalter, and J.D. Cassady, Direct Sequencing with the Aid of Phage Promoters. V.C. Sheffield, D.R. Cox, and R.M. Myers, Identifying DNA Polymorphisms by Denaturing Gradient Gel Electrophoresis. H. Ochman, M.M. Medhora, D. Garza, and D.L. Hartl, Amplification of Flanking Sequences by Inverse PCR. M.A. Frohman and G.R. Martin, Detection of Homologous Recombinants. L.M. Powell, RNA Processing: Apo-B. T.R. Gingeras, G.R. Davis, K.M. Whitfield, H.L. Chappelle, L.J. DiMichele, and D.Y. Kwoh, A Transcription-Based Amplification System. K.D. Friedman, N.L. Rosen, P.J. Newman, and R.R. Montgomery, Screening of ~glgt11 Libraries. Genetics and Evolution. H.A. Erlich and T.L. Bugawan, HLA DNA Typing. J.S. Chamberlain, R.A. Gibbs, J.E. Ranier, and C.T. Caskey, Multiplex PCR for the Diagnosis of Duchenne Muscular Dystrophy. S.B. Lee and J.W. Taylor, Isolation of DNA from Fungal Mtcelia and Single Spores. S.C. Kogan and J. Gitschier, Genetic Prediction of Hemophilia A. U. Gyllensten, Haplotype Analysis from Single Sperm or Diploid Cells. M.L. Sogin, Amplification of Ribosomal RNA Genes for Molecular Evolution Studies. T.J. White, T. Bruns, S. Lee, and J. Taylor, Amplification and Direct Sequencing of Fungal Ribosomal RNA Genes for Phylogenetics. Diagnostics and Forensics. G.D. Ehrlich, S. Greenberg, and M.A. Abbott, Detection of Human T-Cell Lymphoma/Leukemia Viruses. D.E. Kellogg and S. Kwok, Detection of Human Immunodeficiency Virus. I. Baginski, A. Ferrie, R. Watson, and D. Mack, Detection of Hepatitis B Virus. Y. Ting and M.M. Manos, Detection and Typing of Genital Human Papillomaviruses. D. Shibata, Detection of Human Cytomegalovirus. H.A. Rotbart, PCR Amplification of Enteroviruses. D. Mack, O.-S. Kwon, and F. Faloona, Novel Viruses. J. Lyons, Analysis of ras Gene Point Mutations by PCR and Olgonucleotide Hybridization. M. Crescenzi, B-Cell Lymphoma: t(14 18) Chromosome Rearrangement. R.M. Atlas and A.K. Bej, Detecting Bacterial Pathogens in Environmental Water Samples by Using PCR and Gene Probes. S.-H. Park, PCR in the Diagnosis of Retinoblastoma. C. Orrego and M.C. King, Determination of Familial Relationships. Instrumentation and Supplies: R. Watson, PCR in a Teacup A Simple and Inexpensive Method for Thermocycling PCRs. P. Denton and H. Reisner, A Low-Cost Air-Driven Cycling Oven. N.C.P. Cross, N.S. Foulkes, D. Chappel, J. McDonnell, and L. Luzzatto, Modification of a Histokinette for Use as an Automated PCR Machine. C. Orrego, Organizing a Laboratory for PCR Work. R. Madej and S. Scharf, Basic Equipment and Supplies. Index.

Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA

Abstract: We describe a new molecular approach to analyzing the genetic diversity of complex microbial populations. This technique is based on the separation of polymerase chain reaction-amplified fragments of genes coding for 16S rRNA, all the same length, by denaturing gradient gel electrophoresis (DGGE). DGGE analysis of different microbial communities demonstrated the presence of up to 10 distinguishable bands in the separation pattern, which were most likely derived from as many different species constituting these populations, and thereby generated a DGGE profile of the populations. We showed that it is possible to identify constituents which represent only 1% of the total population. With an oligonucleotide probe specific for the V3 region of 16S rRNA of sulfate-reducing bacteria, particular DNA fragments from some of the microbial populations could be identified by hybridization analysis. Analysis of the genomic DNA from a bacterial biofilm grown under aerobic conditions suggests that sulfate-reducing bacteria, despite their anaerobicity, were present in this environment. The results we obtained demonstrate that this technique will contribute to our understanding of the genetic diversity of uncharacterized microbial populations.

Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia.

Abstract: Two new methods were used to establish a rapid and highly sensitive prenatal diagnostic test for sickle cell anemia. The first involves the primer-mediated enzymatic amplification of specific beta-globin target sequences in genomic DNA, resulting in the exponential increase (220,000 times) of target DNA copies. In the second technique, the presence of the beta A and beta S alleles is determined by restriction endonuclease digestion of an end-labeled oligonucleotide probe hybridized in solution to the amplified beta-globin sequences. The beta-globin genotype can be determined in less than 1 day on samples containing significantly less than 1 microgram of genomic DNA.

Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA.

Abstract: Overlapping complementary DNA clones were isolated from epithelial cell libraries with a genomic DNA segment containing a portion of the putative cystic fibrosis (CF) locus, which is on chromosome 7 Transcripts, approximately 6500 nucleotides in size, were detectable in the tissues affected in patients with CF The predicted protein consists of two similar motifs, each with (i) a domain having properties consistent with membrane association and (ii) a domain believed to be involved in ATP (adenosine triphosphate) binding A deletion of three base pairs that results in the omission of a phenylalanine residue at the center of the first predicted nucleotide-binding domain was detected in CF patients

Rapid and simple method for purification of nucleic acids.

Abstract: We have developed a simple, rapid, and reliable protocol for the small-scale purification of DNA and RNA from, e.g., human serum and urine. The method is based on the lysing and nuclease-inactivating properties of the chaotropic agent guanidinium thiocyanate together with the nucleic acid-binding properties of silica particles or diatoms in the presence of this agent. By using size-fractionated silica particles, nucleic acids (covalently closed circular, relaxed circular, and linear double-stranded DNA; single-stranded DNA; and rRNA) could be purified from 12 different specimens in less than 1 h and were recovered in the initial reaction vessel. Purified DNA (although significantly sheared) was a good substrate for restriction endonucleases and DNA ligase and was recovered with high yields (usually over 50%) from the picogram to the microgram level. Copurified rRNA was recovered almost undegraded. Substituting size-fractionated silica particles for diatoms (the fossilized cell walls of unicellular algae) allowed for the purification of microgram amounts of genomic DNA, plasmid DNA, and rRNA from cell-rich sources, as exemplified for pathogenic gram-negative bacteria. In this paper, we show representative experiments illustrating some characteristics of the procedure which may have wide application in clinical microbiology.