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RFLP PCR no amplification
Posted by: rkk_1967 (IP Hidden, New member, 2)
Date: December 21, 2005 08:06AM

Dear Al,
I'm trying to amplify a conserved region of DNA from plant source. But I could not amplify. Only the primers are visible in the gel. Can anyone suggest me to solve this problem. (The buffers, DNA and Taq are working nice with a random primer for RAPD work.)
With Thanks
Kannan

 

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Re: RFLP PCR no amplification
Posted by: kum81 (IP Hidden, Junior member, 10)
Date: December 25, 2005 04:28AM

hi
First of all check that your DNA is all right. Also use Fresh MQ.
2. check that the primers you have designed are right
3. Increase the amount of DNA in the PCR mix
4. put a gradient PCR th find optimum annealing temp
5. Increase elongation time and cycles.
use enhancers like MgCl2, DMSO etc.


hope it solves your problem

best wishes

Kum

 

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