ArticleProceedings of the National Academy of Sciences of the United States of America1994
Effective amplification of long targets from cloned inserts and human genomic DNA.
Article in Proceedings of the National Academy of Sciences of the United States of America, 1994. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 144 papers.
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Who cites it
144 citing papers in PubMed.
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- Genomic characterization of mumps viruses from a large-scale mumps outbreak in Arkansas, 2016.Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases · 2019Article
- Genetic Engineering by DNA Recombineering.Current protocols in chemical biology · 2019Article
- Analysis of FOXO3 Gene Polymorphisms Associated with Human Longevity.Methods in molecular biology (Clifton, N.J.) · 2019Article
- A multiplex reverse transcription PCR and automated electronic microarray assay for detection and differentiation of seven viruses affecting swine.Transboundary and emerging diseases · 2018Article
- FOXO3 longevity interactome on chromosome 6.Aging cell · 2017Article
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- Kinetics of DNA elongation.Doklady. Biochemistry and biophysics · 2017Article
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- Guanine-rich sequences inhibit proofreading DNA polymerases.Scientific reports · 2016Article
- λ Recombination and Recombineering.EcoSal Plus · 2016Review
- Rapid and Accurate Determination of Lipopolysaccharide O-Antigen Types in Klebsiella pneumoniae with a Novel PCR-Based O-Genotyping Method.Journal of clinical microbiology · 2016Article
84 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We have used the polymerase chain reaction (PCR) to amplify up to 22 kb of the beta-globin gene cluster from human genomic DNA and up to 42 kb from phaga lambda DNA. We have also amplified 91 human genomic inserts of 9-23 kb directly from recombinant lambda plaques. To do this, we increased pH, added glycerol and dimethyl sulfoxide, decreased denaturation times, increased extension times, and used a secondary thermostable DNA polymerase that possesses a 3'-to 5'-exonuclease, or "proofreading," activity. Our "long PCR" protocols maintain the specificity required for targets in genomic DNA by using lower levels of polymerase and temperature and salt conditions for specific primer annealing. The ability to amplify DNA sequences of 10-40 kb will bring the speed and simplicity of PCR to genomic mapping and sequencing and facilitate studies in molecular genetics.
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