Hello everybody,
I need to do quantitative PCR/RTPCR (not reverse transcription PCR) for the expression analysis of treated and control explants..I heard that before doing this i need to quantify the cDNA concentration..is this really needed?
kindly explain me in detailed how to do differential expression analysis using RT PCR and what are the requirements?
Dear Srinath,
of course it's crucial to use the same amount of cDNA in all the samples, since you want to get a quantitative output. I guess you use RNA from your samples, reverse transcribe it into cDNA and use this as target? Then you have to measure your RNA samples and transcribe the exact same amount. I don't know if it's also suitable to quantify the cDNA. I have never heard of someone doing this.
I am very much thankful to your answers @Jochen Wilhelm and @Ekaterina Chesnokova.
@Jochen Wilhelm: Yeah Jochen..I am also looking for an workshop on RT-PCR. People always confuses me with different names such as qRT-PCR, reverse trascriptase-PCR, q-PCR.
@Ekaterina: Thank you very much for the information you have provided through the links.
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