20.109(S17):Examine transcript levels in response to DNA damage (Day 4)
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Contents
Introduction
Real-time reverse transcription-polymerase chain reaction (RRT-PCR) allows researchers to monitor the results of PCR as amplification is occurring (this technique is also referred to as quantitative or real-time PCR). During RRT-PCR data are collected throughout the amplification process using a fluorescent dye. The fluorescent dye is highly specific for double-stranded DNA and when bound to DNA molecules the fluorescence intensity increases proportionate to the increase in double-stranded product. In contrast, the data for traditional PCR are simply observed as a band on a gel (remember back to M1D5).
Protocols
Part 1: Setup quantitative PCR assay
Part 2: Discussion of Research article details
Reagents
Next day: Journal club I
Previous day: Assess cell survival and harvest RNA for quantitative PCR assay