smearing in PCR amplification run

Virash Gupta via methods%40net.bio.net (by virashkgupta At gmail.com)
Tue Nov 14 12:26:14 EST 2006


Dear zarrin eshaghi,
I have faced this problem occasionally both using DNA template and PCR
product as template. More often this is due to high protein content in
the DNA product to be run. The proteins are contributed by poor
quality of taq, poor quality of template DNA and sometimes due to high
salt concentration (MgCl2, template DNA). In your case reamplifying
the PCR product did not solve the problem. It seems to be related to
poor quality Taq ( from supplier or home made taq). Try to change the
source of Taq and problem is likely to be solved. Also try using lower
amount of taq per reation as taq itself being protein contributes to
proteins. good luck.

Dr V K Gupta
Insect Molecular Biology lab
Dept of Entomology
Punjab Agricultural University
Ludhiana-Pb-141004-India
virashkgupta At gmail.com



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