Abstract

PCR (polymerase chain reaction) is the most important technique in molecular biology. It allows fast and efficient amplification of nucleic acid sequences and isolation of any target sequence. However, evaluation and analysis of PCR kinetics was less reliable and involved time-consuming methods. New instrumentation for kinetic PCR allows efficient real-time monitoring and quantification of nucleic acid amplification. Accurate DNA quantification within a wide dynamic range can be achieved by monitoring dye-alone or sequence-specific probe generated fluorescence. A simple, economical dye-alone approach has many potential genomic applications, including mRNA quantification for gene expression analysis. It can also be used to confirm the specificity of target sequence with Tm (melting temperature) analysis.

Keywords

Polymerase chain reaction, Post-PCR analysis, Real-time detection of PCR products, Quantitative PCR, Fluorescence energy transfer.