Journal ArticleDOI
Recent advances in the polymerase chain reaction
TLDR
Progresses ranging from the identification of novel genes and pathogens to the quantitation of characterized nucleotide sequences and some recent developments in instrumentation, methodology, and applications of the PCR are presented.Abstract:
The polymerase chain reaction (PCR) has dramatically altered how molecular studies are conducted as well as what questions can be asked. In addition to simplifying molecular tasks typically carried out with the use of recombinant DNA technology, PCR has allowed a spectrum of advances ranging from the identification of novel genes and pathogens to the quantitation of characterized nucleotide sequences. PCR can provide insights into the intricacies of single cells as well as the evolution of species. Some recent developments in instrumentation, methodology, and applications of the PCR are presented in this review.read more
Citations
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Journal ArticleDOI
Inhibition and facilitation of nucleic acid amplification.
TL;DR: This review discusses the findings of many studies related to clinical, food, and environmental microbiology, including approaches that have been used to overcome inhibition and facilitate amplification for detection and typing and describes inhibitors and methods that can overcome the attenuation of amplification.
Journal ArticleDOI
Product Differentiation by Analysis of DNA Melting Curves during the Polymerase Chain Reaction
TL;DR: Analysis of melting curves can extend the dynamic range of initial template quantification when amplification is monitored with double-stranded DNA specific dyes.
Journal ArticleDOI
Bias in Template-to-Product Ratios in Multitemplate PCR
TL;DR: In this paper, the authors explore potential causes and the extent of bias in PCR amplification of 16S ribosomal DNAs (rDNAs), genomic DNAs of two closely and one distantly related bacterial species were mixed and amplified with universal, degenerate primers.
Journal ArticleDOI
A role for hepatitis C virus infection in type II cryoglobulinemia.
TL;DR: Type II cryoglobulinemia is strongly associated with concomitant HCV infection and a high rate of false negative serologic tests, suggesting a role for HCV in the pathogenesis of mixed cryoglOBulinemi.
Journal ArticleDOI
Quantitative RT-PCR: pitfalls and potential.
TL;DR: This review addresses the mathematics of RT-PCR, choice of RNA standards, and quantification strategies (competitive, noncompetitive and kinetic [real-time] amplification) and practical considerations in experimental design.
References
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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.
Journal ArticleDOI
DNA polymorphisms amplified by arbitrary primers are useful as genetic markers
TL;DR: A new DNA polymorphism assay based on the amplification of random DNA segments with single primers of arbitrary nucleotide sequence is described, suggesting that these polymorphisms be called RAPD markers, after Random Amplified Polymorphic DNA.
Journal ArticleDOI
Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia.
Randall Keichi Saiki,Stephen J. Scharf,Fred A. Faloona,Kary B. Mullis,Glenn Thomas Horn,Henry A. Erlich,Norman Arnheim +6 more
TL;DR: Two new methods were used to establish a rapid and highly sensitive prenatal diagnostic test for sickle cell anemia, using primer-mediated enzymatic amplification of specific beta-globin target sequences in genomic DNA, resulting in the exponential increase of target DNA copies.
Book ChapterDOI
Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction.
Kary B. Mullis,Fred A. Faloona +1 more
TL;DR: A method whereby a nucleic acid sequence can be exponentially amplified in vitro is described in the chapter, and the possibility of utilizing a heat-stable DNA polymerase is explored so as to avoid the need for addition of new enzyme after each cycle of thermal denaturation.
Journal ArticleDOI
Chelex 100 as a Medium for Simple Extraction of DNA for PCR-Based Typing from Forensic Material
Walsh Ps,Metzger Da,Higuchi R +2 more
TL;DR: The extraction of DNA from semen and very small bloodstains using Chelex 100 is as efficient or more efficient than using proteinase K and phenol-chloroform extraction and DNA extracted from bloodstain seems less prone to contain PCR inhibitors when prepared by this method.