cDNA -mRNA hybrid - (Jun/28/2006 )

Dear everybody
what the NaOH traitement to separate cDNA-mRNA hybrid after cDNA first strand synthesis.

why?

what goal are you trying to accomplish?

I wont to polyA tail first strand cDNA with terminal transferase and using oligidT for the second strand synthesis and for PCR amplification of the cDNA pool.

hmmm

najib, I have not added a polyA tail and so cannot answer that question

however, if you want to do PCR on the hybrids, I think they would probably work just fine as they are? or, use an RNase treatment

oh, and I just saw your PM. please note that I am not here all day every day; I certainly haven't been ignoring you

Can't you use an RnasH positive Reverse transcriptase? They should give you single stranded cDNA and destroy the RNA durin RT?