LONG PCR. HELP

Blanca Barquera barquera at rock.helsinki.fi
Mon Dec 4 06:45:50 EST 1995


I have tried to amplify a 15 kb fragment from chromosomal DNA 
from Paracoccus denitrifcans, using "Expand long template PCR 
system from Boehringer. I have tried several conditions and  I 
have gotten nothing or a big product (20kb) unrelated with what I 
want. I have used DMSO to denature DNA also unsucesfully. My DNA 
is rich in G+C.I have changed buffers, template concentration (50 
to 250 ng. Mg concentration and amount of enzyme. I do not know 
what to change.My primers have worked to get smaller fragments.
Some help would be appreciated. Thank you
Blanca Barquera
University of Helsinki
 




More information about the Methods mailing list