PCR in unknown samples

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PCR in unknown samples

Post by ssas » Fri Aug 18, 2006 5:02 pm

When doing PCR, primers of the target segment of interest have to be synthesized and are necessary for the reaction, right?

How come if we have to amplify DNA from little-known samples or from samples of ancient animals which of course their base sequence is still unclarified. How can we create primers to be used in such cases?

Thank you in advance for your kind suggestions.

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Inland Taipan
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Post by canalon » Fri Aug 18, 2006 6:05 pm

If you are not sure of the sequence, the best is to go with long primers in a conserved region, and used degenerate primers (when ordering your primers, you can ask some base to be replaced by any purine, pyrimidine or the four bases or any combination).

Science has proof without any certainty. Creationists have certainty without
any proof. (Ashley Montague)

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