Target amplification for DNA analysis by the polymerase chain reaction. 1990

K B Mullis

The polymerase chain reaction (PCR) has become a standard laboratory technique. An enzymatic reaction, as simple to perform as it is satisfying to contemplate, the PCR solves two of the more universal problems in the chemistry of natural nucleic acids. It allows for the physical separation any particular sequence of interest from its context; and then provides for an in vitro amplification of this sequence which is virtually without limit. The surprising robustness of PCR derives from its fortuitous combination of three phenomena, each of which is intrinsically powerful. The first of these is the impressive ability of almost all oligodeoxynucleotides to bind tightly and specifically to their complementary nucleic acid sequences, discriminating easily between hundreds of thousands of sites. The second familiar phenomenon is illustrated by the notion that the probability for the occurrence of a compound action is the product of the individual probabilities for the occurrence of each of its components. The third phenomenon embodied in the polymerase chain reaction relates to the branching structure of its propagation and the inherent robustness attached to such a form. Consideration of the above leads to certain generalities regarding the relative utility of various protocols for carrying out the PCR. Specific conditions of time, temperatures, concentrations, etc. will be described, as well as sample preparation and analytical methods.

UI MeSH Term Description Entries
D013995 Time The dimension of the physical universe which, at a given place, orders the sequence of events. (McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed) Effects, Long-Term,Effects, Longterm,Long-Term Effects,Longterm Effects,Effect, Long-Term,Effect, Longterm,Effects, Long Term,Long Term Effects,Long-Term Effect,Longterm Effect
D015342 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. Chromosomal Probes,DNA Hybridization Probe,DNA Probe,Gene Probes, DNA,Conserved Gene Probes,DNA Hybridization Probes,Whole Chromosomal Probes,Whole Genomic DNA Probes,Chromosomal Probes, Whole,DNA Gene Probes,Gene Probes, Conserved,Hybridization Probe, DNA,Hybridization Probes, DNA,Probe, DNA,Probe, DNA Hybridization,Probes, Chromosomal,Probes, Conserved Gene,Probes, DNA,Probes, DNA Gene,Probes, DNA Hybridization,Probes, Whole Chromosomal
D016133 Polymerase Chain Reaction In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships. Anchored PCR,Inverse PCR,Nested PCR,PCR,Anchored Polymerase Chain Reaction,Inverse Polymerase Chain Reaction,Nested Polymerase Chain Reaction,PCR, Anchored,PCR, Inverse,PCR, Nested,Polymerase Chain Reactions,Reaction, Polymerase Chain,Reactions, Polymerase Chain

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