can I use routine PCR to assess whether a gene is expressed or not?
Posted 26 April 2010 - 12:50 AM
I designed a pair of specific primers for this gene using a RT-PCR primer design software, Beacon Designer. So can I use cDNA as template and the pair of specific primers to run a routine PCR, and if there is band with correct length after electrophoresis, I can conclude that this gene is expressed?
Thanks a lot.
Posted 26 April 2010 - 03:10 AM
Posted 26 April 2010 - 03:43 AM
Posted 26 April 2010 - 08:15 AM
Transcription is a fuzzy process, not ON/OFF like a switch. Therefore, if you have enough cells, you'll get a band for everything, inluding non-transcribed genes. I'd recommend to use a positive and a negative control. Then you can say: compared to gene A, which is usually not transcribed, and similar to gene B, which is usually transcribed... So you'll end up with some kind of transcription level.