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Old 01-13-2010, 06:35 AM   #13
Anri
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Location: Estonia

Join Date: Jan 2010
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Quote:
Originally Posted by bmcjc View Post
I want to sequence my PCR product on Solexa (it's a long story). Will it be feasible to PCR amplify with primers that has the Illumina's long PCR primer sequence extended to the 5'end so I don't have to do adaptor ligation again?


Check this article. Is anyone done something like this? (http://www.ncbi.nlm.nih.gov/pmc/arti...ukmss-3213.pdf).

Direct sequencing of short amplicons
To avoid unnecessary PCR amplification steps, which would potentially exacerbate biases, we can perform extremely deep sequencing of short amplicons using locus-specific primers that possess tails that are capable of hybridisation to the oligos tethered to the flowcell surface. The tailless forward and reverse oligos are then used as primers in the sequencing steps
(Supplementary Protocol 10 online).
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