la pcr complexes

Shahram Mori smori at nmsu.edu
Sat Mar 11 22:58:28 EST 1995


aEscher wrote:
: We are amplifying a 6 kb fragment from mouse genomic DNA. No problem, but
: suddenly from one day to the next all we can see is a high MW complex 
: stuck in the well. This phenomenon seems to be the result of the amplifi-
: cation reaction. Does anyone know what's happening? 
: Thanks,
: Alan Escher.
:                              

Is it possible that prior to loading, the DNA wasn't heated up? 
Especially with large DNA's the folding of DNA on itself my prevent it 
from entering the gel. I usually place my DNA at 65C (10min) and quicly chill
on ice prior to loading.
Cheers
--
Shahram Mori					   _/\_
Program in Molecular Biology			  _\  /_
Dept. of chemistry and Biochemistry Box 3C	  \_  _/
NMSU  Las Cruces NM				    ||
88003





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