Real Time PCR problem - different dilutions have the same Ct
Posted 29 April 2004 - 07:33 AM
Posted 29 April 2004 - 11:08 AM
Contamination is a pain to eliminate. Even thorough cleaning of all pipettes, replacement of all reagents, cleaning bench tops, etc. sometimes fails to eliminate the source.
Change all reagents (again)
Setup your reaction using a different labs pipettes and workspace (different as in ANOTHER ROOM!)
Oh, are you using TaqMan or SYBR Green? What C(T) are things coming up at? Also, is it for just one assay or all assays?
Edited by unknownphd, 29 April 2004 - 11:09 AM.
Posted 29 April 2004 - 05:21 PM
Posted 04 March 2005 - 06:52 AM
Posted 07 March 2005 - 03:45 AM
However, if you are using TaqMan, I guess you've still got contamination, as suggested before. The only other option, if you're sure that is not contamination, is that your probe is somehow degrading, but I can not think of a reason for this to happen. Maybe you could run your samples in a gel after the PCR, specially the blank, to determine if it is or not contamination.
Hope this helps!