Hello,
is there anyone studying on HIV proteins? I'm studying on rev and tat. I have few problems and need specific answers.

You can post your questions here and say if anyone can answer you.
Tracy

First, I'm trying to clone my inserts tat and rev into pJET1/blunt vector. It is a positive selection vector. You can use non-purified PCR products in ligation reaction and after transformation you plate the mixture LB amp plates. You don't need to use XGal or IPTG. Due to a lethal gene, only recombinant colonies survive. I got at least 30 colonies in a plate. Although I easily cloned tat, I couldn't find a plasmid including rev insert. (I did colony PCR to all colonies). It is really interesting. I repeated it with purified PCR product, but the result is same. Is there any idea why colonies appear but have no insert?

Have you tried other templates to PCR from?

I have already overcome my problems, thanks dvatakis. By the way, I used other templates.