RT PCR Standard curve amplification problem - (Oct/15/2007 )
Hi folks....I am having a problem amplifying my real time RT PCR standards. I am doing an absolute quantification curve using ds DNA. These standards have amplified pretty well in the past (a few weeks ago!) and now they didnt amplify ![]() The good news is my unknowns amplified on the same plate. I use the same mastermix, the same detectors, and of course the same primers and probes for my protein marker. I am quite confounded as to what is going on with my standards. Surely they couldn't have degraded, I mean all 10 of them?! Seems quite impossible, but I really dont know what else to suspect?! Any thoughts on what could be going on?
 The good news is my unknowns amplified on the same plate. I use the same mastermix, the same detectors, and of course the same primers and probes for my protein marker. I am quite confounded as to what is going on with my standards. Surely they couldn't have degraded, I mean all 10 of them?! Seems quite impossible, but I really dont know what else to suspect?! Any thoughts on what could be going on? 
Thank you ![]()
-kaya-
Are you sure that the heating block of your cycler is alright?
-krümelmonster-
