Subtitle:

Real Time PCR. The idea of the method and strategies of reaction monitoring

Publisher:

Committee on Biotechnology PAS ; Institute of Bioorganic Chemistry PAS

Abstract:

Real-time PCR has become one of the most widely used methods of gene quantitation in molecular biology and medical diagnostics. This technique combines PCR amplification and the detection of the PCR product into a single step. In real-time PCR, the amount of product formed is measured during the course of the experiment by monitoring the fluorescence of dyes or probes introduced into the reaction. Fluorescence data are generated by the use of intercalating dyes such as SYBR Green 1 or molecular probes, the most important of which are: TaqMan, Molecular Beacons, Hybridization Probes, and Scorpion Probes. The real-time PCR reactions are characterized by the PCR cycle in which the target amplification is first detected. This cycle is referred to as a treshold cycle (Ct), at which fluorescence intensity becomes greater than background fluorescence. Consequently, the greater the quantity of target DNA in the starting material, the faster the significant increase in fluorescence intensity will appear, yielding lower Cf The relation between Ct and the concentration of the target sequence allows for a precise quantitation of the genetic material in the sample.

Relation:

Biotechnologia, vol.80, 1 (2008)-.

Volume:

80

Issue:

1

Start page:

71

End page:

85

Detailed Resource Type:

Article

Format:

application/pdf

Resource Identifier:

0860-7796 ; oai:rcin.org.pl:74061

Source:

Library of Institute of Bioorganic Chemistry PAS

Language:

pol

Language of abstract:

eng

Temporal coverage:

1988-2010

Rights:

Creative Commons Attribution BY-SA 4.0 license

Terms of use:

Copyright-protected material. [CC BY-SA 4.0] May be used within the scope specified in Creative Commons Attribution BY-SA 4.0 license, full text available at:

Digitizing institution:

Institute of Bioorganic Chemistry of the Polish Academy of Science

Original in:

Institute of Bioorganic Chemistry of the Polish Academy of Science

Projects co-financed by:

Operational Program Digital Poland, 2014-2020, Measure 2.3: Digital accessibility and usefulness of public sector information; funds from the European Regional Development Fund and national co-financing from the state budget.

Access:

Open