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Old 09-27-2009, 06:02 PM   #11
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Location: Lexington KY

Join Date: Jun 2009
Posts: 4

My sequences were on ncbi so I just used it to design primers.

Or you could find highly conserved regions, create primers, sequence, and repeat until you get the whole sequence.

I believe this step is imperative. If you are using Poly A purification then any Rnase contamination or degradation before the last purification step will lead to sequences further from the A tail not being incorporated and thus not sequenced.

This is why we designed primers around the 5' end of the cdna.
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