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Author Topic:   Difficulty in PCR amplification
posted 09-24-2000 01:45 PM     Click Here to See the Profile for Administrator     Edit/Delete Message Reply w/Quote
Dear Dr

In a RAP-PCR experiment with Chinese hamster cell line under two different environmental conditions we obtained avery good fingerprinting pattern. however, during reamplification we faced a problem. The elution and reamplification has been done before in our lab. I am a little puzzled why it is not working. We guess something is wrong with the primer. Can it happen like that? DNA concentration is not a problem because the same primer is not working with other genomic DNAs as well. Primer seq is 5' GACCATTGCATAACGCTGAC 3' Several PCR conditions were tried with Boringer Manheim Taq and the buffer supplied.The same Taq worked with other samples. Can you suggest something of help?

Thanking you

Sincerely yours

Dr Keya Chaudhuri


posted 06-06-2004 12:01 AM     Click Here to See the Profile for daforerog   Click Here to Email daforerog     Edit/Delete Message Reply w/Quote
Try PCR adjuvants such as BSA or DMSO. It may be useful.

Good luck.



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