Quantification of mRNA and analysis of mutation by capillary PCR.

If I classify roughly for the quantification method of mRNA, there are the following 5 kinds. 1. Northern and dot hybridization, 2. RNase protection assay, 3. RT-PCR (the use of internal control), 4. competitive RT-PCR (the use of competitor), 5. real time monitoring PCR. In these methods, 3-5 employ PCR. Though 3, 4 are a method to quantify at an exponential increase term, it is different point that 5 is quantification method by means of PCR cycle number to exceed a detection limit of PCR product, just before entering an exponential increase term. Recently, a quantification method by the real time monitoring PCR basks in attention. Not only this method isn't necessary to confirm a cycle number of an exponential increase term in advance but have the wide quantification range in comparison to the method to quantify at an exponential increase term, there are many merits. I introduce LightCyclerTM system (Roche Diagnostics) with this draft as an equipment to be able to do a realtime monitoring.