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Old 02-26-2011, 12:36 PM   #1
zhengz
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Location: sweden

Join Date: Aug 2010
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Default read length limits sample prep time?

It seems very critical to increase PCR time to generate less biased long amplicons such as >200, >400, >1kb? Trying to reduce sample prep time including emPCR might, at least currently, be limited by a sufficient time for emPCR.

Not only accuracy and length, GC content is another issue. The recent Broad paper (on Illunima library prep) using increased time for the 98c step, from seconds to >1 min, plus some other adjustments, improved results with less GC biased reads.
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