Protocol Online logo
Top : Forum Archives: : Molecular Biology

Long PCR problem - Product smaller than expected size (Jan/29/2003 )

I have been using the Roche Expand Long PCR kit (system 3) and keep getting a product about half my expected size (`9000bp vs ~15000bp); I am trying to amplify the entire mitochondrial genome of Orthopteran insects. In the course of my experiments, I also often get what appear to be very large pieces of DNA hung up in the wells of theagarose gels that I run to screen my products. Any ideas?

-lsenior-

hai
if u have long well platform to run ur amplified products then use it
size of the well should be 5 cm .
i also had the same probs but then i used 5cm well and solved
bye

-nanibio-

You might want to try software designed for picking primers for long PCR or high-low GC contents:

http://www.changbioscience.com/primo/primohl.html

Chang

-czhu-

I also had the same problems. Has your problem been rectified? If yes, please let me know the key. Thanks.

-vgupta-