cDNA library screening with 5' specific primer - (Mar/31/2006 )
i have to do cDNA library screening by using 5' specific primers for a particular gene.
how efficient it is when compared to 3' specific primer and using both the primers?will it be efiicient if my gene of interest is rare in the library?
please suggest me how to proceed.
thank you in advance
If your 5' primer a reverse one and 3' primer is a forward gene specific primer, I'd feel you should use bother.
You could not guarantee intact 5’ end of cDNA products in library even though you might prepare the library by RLM or SMART strategy. Using 5’ reverse primer with 5' adaptor primer or primer on vector might easily obtain smear signal, especially for cDNA library prepared by polyT primer for 1st strand cDNA…