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Re-amplification of PCR product
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Re-amplification of PCR product
I try to re-amplify the product with the same first round pcr primers.
Although I purified the first round pcr product with different method.
Instead ofobtaining strong amplfied band I get weak band with lots of non-specific bands ... ...
Last update 26-Apr-2002, Rating Good of 0 votes.
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to cut the long story short, i was getting bacterial contamination while trying to pcr a plant based two component system which has the same conserved regions as bacterial. Bactera elimination from the roots is impossible. The degenerate primers are pcking up the bacterial two component sytem.
I then decided to to amplify from the genomic DNA with specific primers, that worked fine. Cleaned the product and made series of dilutions. Tried to to amplify using the degenerate primers and no success. Opitimized other parameters still no sucess. HELP. thaks hope to hear from you soon
Rating: Good
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what are the reasons for these dilution?
Thanks
Rating: Good
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Maybe you should try diluting your PCR products before reamplification. You can tru several dilutions and see which one gives the best result.
Rating: Good
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