PCR1

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PCR1

polymerase chain reaction. The amplification of a specific DNA sequence, termed target or template sequence, that is present in a complex mixture, by adding two or more short oligonucleotides, also called primers, that are specific for the terminal or outer limits of the template sequence. The template-primers mixture is subjected to repeated cycles of heating to separate (melt) the double-stranded DNA and cooling in the presence of nucleotides and DNA polymerase such that the template sequence is copied at each cycle. Thermostable polymerases such as those obtained from a hot springs bacterium Thermus aquaticus, commonly termed Taq polymerase, are used.
At the end of 20 to 30 such cycles, the amplified target sequence, which may have been present in as few as a single copy in the original mixture, can be readily detected, for example by electrophoresis and ethidium bromide staining in an argarose gel.

multiplex PCR
the detection of more than one template in a mixture by addition of more than one set of oligonucleotide primers.
nested PCR
the primers used in the first round of amplification are either both replaced (nested PCR) or only one is replaced (semi-nested PCR) for the second and subsequent cycles of amplification. Increases the sensitivity and specificity of the PCR.
quantitative PCR
a means for quantifying the amount of template DNA present in the original mixture. Usually achieved by the addition of a known amount of a target sequence that is amplified by the same primer set but can be differentiated, usually by size, at the end of the reaction.
real-time PCR
a method for the detection and quantitation of an amplified PCR product based on incorporation of a fluorescent reporter dye; the fluorescent signal increases in direct proportion to the amount of PCR product produced and is monitored at each cycle, 'in real time', such that the time point at which the first significant increase in the amount of PCR product correlates with the initial amount of target template.
reverse-transcriptase PCR (RT-PCR)
a reaction applied when the target sequence is RNA, such as viral RNA or messenger RNA. Reverse transcriptase that copies DNA from an RNA template is present in the first round.
References in periodicals archive ?
Additionally, the ResQ enables sensitive and accurate quantification of target RNAs by real-time quantitative PCR (RT-qPCR) using amplicons that are much shorter (down to 20 bp) than is typically required for RT-qPCR methods, said the company.
Quantitative PCR monitors the amplification of the specific DNA during the polymerase chain reaction extension time.
On this account, we used real-time quantitative PCR (probe method) to detect content of BCV and JCV in renal transplant recipients, aiming to provide a valuable basis for clinical diagnosis and therapeutic schedule.
That's why scientists developed a quantitative PCR method for quantifying individual strains in a commercial yogurt starter culture.
Then to verify their expression, he used real time quantitative PCR and immunostaining with antibodies.
Real-time quantitative PCR (qPCR) is an advanced technology for quantification of nucleic acids, gene expression studies and has become an essential need in diagnostic research works.
We are in the process of expanding our Molecular Infectious Disease testing for our Nephrologists and Infectious Disease docs to perform the quantitative PCR for HBV, HCV and HIV.
A decade ago, he began using a technique known as quantitative PCR to get definitive answers about how much of a particular life form is present in a sample of wine or juice.
Both simple and easy to use with existing real time quantitative PCR equipment.
Quantification of target sequences via real-time PCR or quantitative PCR (qPCR) is based on the detection and measurement of amplified products (amplicons) during each cycle of the amplification reaction.
The remaining chapters review key areas of current molecular research, such as microarray analysis, quantitative PCR, and transgenics.
Stephen Gunstream, IDT's OEM Business Unit Leader commented, "Our new qPCR scales ensure that all scientists, including those who only require limited reactions, can have access to the most precise quantitative PCR chemistry available.

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