cDNA PCR - no product - questions - HELP!!!!
I have isolated cDNA and it look ok after PCR using housekeeping genes. Anyhow as soon as I try primers (blasted etc) for the gene of intrest there is no or a wrong product... even primer that used to work for genomic PCR seem not to work at all anymore. :unsure::confused:
So anyhow I was wondering what to change, I tried optimizing with temperature or Mg or Q Solution but still nothing. I read reverse transcriptase inhibits Taq. So do you people clean up your product from reverse transcription or use it directly for pcr amplification? Do I need to change PCR conditions if using cDNA instead of genomic DNA? :wacko:
Slowly this starts to become really frustrating. So I would be really THANKFULL if anyone here had any ideas how to improve this...
Re: cDNA PCR - no product - questions - HELP!!!!
|All times are GMT. The time now is 11:39 PM.|
Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved