Real-time PCR about Incorrect setup of the data collection stage - (May/09/2011 )
Recently, I ran real-time PCR for TaqMan MicroRNA assay using ABI 7500, but I set up a incorrect data-collection stage in stage 1 (95 degrees centigrade for 10 minutes,1 cycle).Interestingly, the final result appeared that CT value is about 28. Why is the CT value about 28 in the stage 1 for data collection?
TaqMan probes are cleaved in the elongation stage, and free reporter dyes remain in the reaction to the next denaturation step. IMHO.