it's good to be back!
I have got this problem with a viral genome from which I have to produce cDNA.
I have extracted the total viral nucleic acid and have used the "transcriptor high fidelity cdna synthesis kit" by the grate company: Roche. I expect to get fragments between 3.5 to 4 kb. the problem is that the first 700 nucleotides of the viral genome( which by the way is a ss positive rna genome) is highly structures and gc rich.
there is the dilemma:
do i increase the temperature or do i prolong the incubation time or do i have to do both? is there a best way to reach the balance between these two factors?
I attache the catalogue in case any one decides to take look and help me out:))