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Old 08-02-2017, 10:30 AM   #27
chaomeizhang
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Location: Tampa, Florida

Join Date: Aug 2011
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Quote:
Originally Posted by Chipper View Post
It is because you have two different adaptors (annealed in a Y-shape). Without dUTP you will get two different amplicons for each insert due to the Y-adaptors meaning that the first read can start from either end of the insert, but with dUTP you will only amplify one amplicon (which will be double stranded) so all reads from the same transcript will go in the same direction.
Thank you Chipper.

I would like to add some note to the explanation.

The enrichment PCR does regenerate the DNA strand which was degraded by UDG, but the adaptors on both sides of the strands can not hybridized with the lawn oligos on the flow cell, so these strands will be excluded during the cluster generation and eventually not be sequenced.
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