|Register||Search||Today's Posts||Mark Forums Read|
|PCR - Polymerase Chain Reaction Forum PCR - Polymerase Chain Reaction Forum. Discuss and ask questions about PCR troubleshooting, PCR protocols and methods, PCR products, and PCR theory.|
| ||LinkBack||Thread Tools||Display Modes|
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.
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?
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!!!!
|cdna , pcr , product , questions|
|Thread||Thread Starter||Forum||Replies||Last Post|
|100 Questions for Plant Science Research||Claire Grierson,School Biological Sciences||Arabidopsis and Plant Biology||2||02-11-2013 07:00 AM|
|Product detectible on gel, melting curve peak, but no amplification curve||emucaki||Real-Time PCR and Quantitative PCR Forum||4||08-24-2009 08:54 PM|
|100 Questions Deadline JUST TWO WEEKS AWAY||Claire Grierson,School Biological Sciences||Arabidopsis and Plant Biology||0||03-17-2009 03:52 PM|
|cDNA Synthesis and Amplification of POLY(A) mRNA by RT-PCR Protocol||domba||Real-Time PCR and Quantitative PCR Forum||3||09-09-2008 02:48 PM|