PCR Multiplexing Based on a Single Fluorescent Channel Using Dynamic Melting Curve Analysis.

ABSTRACT: Since its invention in 1986, the polymerase chain reaction (PCR), has become a well-established method for the detection and amplification of deoxyribonucleic acid (DNA) with a specific sequence. Incorporating fluorescent probes, known as TaqMan probes, or DNA intercalating dyes, such as SYBR Green, into the PCR mixture allows real-time monitoring of the reaction progress and extraction of quantitative information. Previously reported real-time PCR product detection using intercalating dyes required melting curve analysis (MCA) to be performed following thermal cycling. Here, we propose a technique to perform dynamic MCA during each thermal cycle, based on a continuous fluorescence monitoring method, providing qualitative and quantitative sample information. We applied the proposed method

SUBMITTER: Zhang H 

PROVIDER: S-EPMC7689941 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature