Journal ArticleDOI
High-fidelity amplification using a thermostable DNA polymerase isolated from Pyrococcus furiosus
Kelly S. Lundberg,Daniel D. Abrahms Apartment C Shoemaker,Michael W. W. Adams,Jay M. Short,Joseph A. Sorge,Eric J. Mathur +5 more
TLDR
These studies demonstrate that the error rate per nucleotide induced in the 182 known detectable sites of the lacI gene was 1.6 x 10(-6) for Pfu DNA polymerase, a greater than tenfold improvement over the 2.0 x 10 (-5) error rate for Taq DNA polymerases.About:
This article is published in Gene.The article was published on 1991-12-01. It has received 685 citations till now. The article focuses on the topics: Pfu DNA polymerase & Hot start PCR.read more
Citations
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Cd-hit: a fast program for clustering and comparing large sets of protein or nucleotide sequences
Weizhong Li,Adam Godzik +1 more
TL;DR: Cd-hit-2d compares two protein datasets and reports similar matches between them; cd- Hit-est clusters a DNA/RNA sequence database and cd- hit-est-2D compares two nucleotide datasets.
Journal ArticleDOI
Determination of microbial diversity in environmental samples: pitfalls of PCR‐based rRNA analysis
TL;DR: Specific aspects of sample collection, cell lysis, nucleic acid extraction, PCR amplification, separation of amplified DNA, application of nucleic probes and data analysis are covered.
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PCR amplification of up to 35-kb DNA with high fidelity and high yield from lambda bacteriophage templates
TL;DR: The base-pair fidelity, the ability to use PCR products as primers, and the maximum yield of target fragment were increased and improvements were achieved by the combination of an exonuclease-free, N-terminal deletion mutant of Taq DNA polymerase, Klentaq1.
Journal ArticleDOI
Developments in industrially important thermostable enzymes: a review.
TL;DR: The source microorganisms and properties of thermostable starch hydrolysing amylases, xylanases, cellulases, chitinases, proteases, lipases and DNA polymerases are discussed and the industrial needs for such specific thermostably enzyme and improvements required to maximize their application in the future are suggested.
Journal ArticleDOI
Transmission of Hepatitis C by Intrahepatic Inoculation with Transcribed RNA
Alexander A. Kolykhalov,Eugene Agapov,Eugene Agapov,Keril J. Blight,Keril J. Blight,Kathleen Mihalik,Kathleen Mihalik,Stephen M. Feinstone,Stephen M. Feinstone,Charles M. Rice,Charles M. Rice +10 more
TL;DR: This work defines the structure of a functional HCV genome RNA and proves that HCV alone is sufficient to cause disease.
References
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A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding
TL;DR: This assay is very reproducible and rapid with the dye binding process virtually complete in approximately 2 min with good color stability for 1 hr with little or no interference from cations such as sodium or potassium nor from carbohydrates such as sucrose.
Journal ArticleDOI
Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase
Randall Keichi Saiki,David H. Gelfand,Susanne Stoffel,Stephen J. Scharf,Russell Higuchi,Glenn Thomas Horn,Kary B. Mullis,Henry A. Erlich +7 more
TL;DR: A thermostable DNA polymerase was used in an in vitro DNA amplification procedure, the polymerase chain reaction, which significantly improves the specificity, yield, sensitivity, and length of products that can be amplified.
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Specific Enzymatic Amplification of DNA In Vitro: The Polymerase Chain Reaction
Kary B. Mullis,Fred A. Faloona,Stephen J. Scharf,Randall Keichi Saiki,Glenn Thomas Horn,Henry A. Erlich +5 more
TL;DR: An alternative method for the synthesis of specific DNA sequences is explored that involves the reciprocal interaction of two oligonucleotides and the DNA polymerase extension products whose synthesis they prime, when they are hybridized to different strands of a DNA template in a relative orientation such that their extension products overlap.
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Fidelity of DNA synthesis by the Thermus aquaticus DNA polymerase.
TL;DR: The fidelity of in vitro DNA synthesis catalyzed at high temperature by the DNA polymerase from the thermophilic bacterium Thermus aquaticus is determined and it is demonstrated that the purified polymerase lacks detectable exonucleolytic proofreading activity.
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Pyrococcus furiosus sp. nov. represents a novel genus of marine heterotrophic archaebacteria growing optimally at 100°C
Gerhard Fiala,Karl O. Stetter +1 more
TL;DR: Ten strains representing a novel genus of marine thermophilic archaebacteria growing at between 70 and 103°C with an optimal growth temperature of 100°C and a doubling time of only 37 min were isolated from geothermally heated marine sediments at the beach of Porto di Levante, Vulcano, Italy.