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Old 03-09-2011, 07:56 PM   #1
RNAseqer
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Default Size selection after Covaris shearing...necessary?

With a show of hands, how many people still do size selection post ligation after shearing genomic DNA by Covaris ? I've heard that many skip size selection now, but many others still size select.
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Old 03-10-2011, 05:49 AM   #2
Heisman
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For exome capture we don't do size selection. We purify with SPRI beads and that adequately removes adapter/primer-dimer.
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Old 03-10-2011, 07:07 AM   #3
greigite
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Depends how tight of a size distribution you want. We size select post-Covaris to get a narrow distribution allowing assembly of overlapping paired-end reads. Size selection can also help you get longer read lengths since all PCR based sequencing approaches (454, Illumina) do tend to enrich for smaller fragments.
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