PCRing mature form of a gene - (Aug/15/2011 )
I recently PCRed my gene of interest (the complete form of the gene) from genomic DNA and now another step is to only amplify the gene encoding for the mature form of my gene and then ligate into an expression vector...how do I ONLY amplify the mature form..wouldn't I essentially do the exact same thing as the first time I did PCR the only diff. being I'll be ligating into an expression vector? And then the mature protein will be produced once in expression vector?? Hope I'm making sense...
I think you did not do the first step right. You should directly amplify the whole gene from cDNA which is converted from RNA by RT reaction.