Amplification of 3.4 kb product from RNA for cloning
Posted 26 December 2010 - 05:02 AM
Posted 26 December 2010 - 06:58 AM
Posted 26 December 2010 - 11:35 PM
So what Tm shd I consider? Inclusive of this extra bases (which the clontech protocol says u dont have to). I really dont know. I am not able to amplify my gene of interest using these primers.
Posted 27 December 2010 - 07:53 AM
Posted 27 December 2010 - 09:57 PM
Posted 05 January 2011 - 05:16 AM
Doing two step RT-PCR I also often had problems to amplify larger fragments (>1 kB) from cDNA. In my opinion the cDNA one gets after reverse transcription is kind of fragmented, making amplification of longer products difficult.