long PCRs: Yes, it works on human DNA !

[Eric Campeau]

Hi everyone,

  I have been using inverse PCRs to determine my gene structure as an easy way to do on the side of other experiments.  I am studying a human gene and for me, 
some of my clones were up to 1.4-1.6 kb  (I subcloned my PCR bands using the TA cloning kit).  I have found about 15 intron/exon junctions by this way.  I did not try to get 
longer PCR products since I was interested only in the surrounding sequences (that is why I used four base cutters like NlaIII, AluI, MseI ... etc).  Anyway, if some of you are 
interested in the protocol we are using, e-mail me or just ask me to post it and I'll do it.  Hope this will help !


Eric Campeau
Graduate student
McGill University
e-mail: popa0272 at po-box.mcgill.ca