RT-PCR reagents KIT 1 or 2-step? - (Jul/10/2008 )
I ma new in the PCR field. We have not PCR jet, first I collect information about equipment, reagents. What I want is RNA isolation from human whole blood, and then analyse it on PCR, and now I have problem, because I don't know which RT-PCR kits is better 1-stpe or 2-step. Could someone give me some advice, and write difference between 1- and2-step PCR reactions? Please help!!
Two-step method gives you a flexible approach, once you finish cDNA step, you can test different genes later one in the future.
Yep I would also go for two step RT-PCR. If you are working with infectious samples (virus detection) it may be more advantegous to use 1-Step.
I would go for SuperScript (Invitrogen but more expensive) or Transcriptor (Roche Applied Science). These 2 kits are really the gold-standard.
Roche offers their new product Transcriptor HiFi as a free sample at the moment. Maybe you want to try it. My collegue got nice results with it. I only worked with the conventional Transcriptor 1st strand synth. kit which is at least as sensitive as SS is...
Thanks for answers.. I will be working with viruses- HIV, HCV and HBV, actually I need equipment and Kits which can be use in diagnostic. I found that Roche has such tests -COBAS TaqMan HCV/HBV/HIV Tests but all is writing that the test is intended for use in conjunction with clinical presentation and other markers as an aid i assessing viral response, and that it is not intended for use as a screening test for the presence of HCV in blood or blood products:(