Real-Time Reverse-Transcription Polymerase Chain Reaction: Technical Considerations for Gene Expression Analysis

摘要: The reverse transcription - polymerase chain reaction (RT-PCR) is a sensitive technique for the quantification of steady-state mRNA levels, particularly in samples with limited quantities extracted RNA, or analysis low level transcripts. procedure amplifies defined transcripts by taking advantage retroviral enzymes transcriptase (RT) activity, coupled to PCR. resultant PCR product concentration directly proportional initial starting quantity mRNA, therefore allowing gene expression incorporation fluorescence detector appropriate amplicons. In this chapter, we describe number most popular techniques performing RT-PCR and detail subsequent methodologies required interpret data either relative manner through absolute levels.