Transcription Rate - Measure Hn RNA using Taqman?? (Oct/01/2004 )
Could someone tell me how the transcription rate can be measured using TaqMan (real-time PCR) ?? If I design primers spanning an exon and an intron, will that give me a measure of the HnRNA present and thereby give me the transcription rate?
to get to the transcription-rate is quite straightforward. you need a rt-pcr for your gene, that spans an exon, so to say only real, processed mRNA wil give a signal, and anotherone for a gene stably transcribed under whatsoever conditions (usually a housekeeping gene like GAPD-H). You do the RT-PCR and the qPCR with both set of primer, you will get transcription rates like 10 copies of your GOI / 1 copy GAPD-H.