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Old 01-16-2014, 11:45 AM   #1
slees
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Location: NC

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Default How to fill the gap after tagmentation? Can I use Klenow fragment?

I am trying to making Illumina sequencing library using transposon based method to add adaptor. However, instead of using the adaptor used in Nexera kits, I would like to use the Y-shaped adaptor. Thus, the gap between the bottom stand of adaptor and bottom strand of genomic DNA need to be repaired. I want to find a method to do gap filling, however, I do not want the DNA to the 3' of the gap (i.e., bottom strand of the adaptor) to be removed by 5'-3' exonuclease activity of DNA Pol I and then re-synthesized. Can I use Klenow fragment here? It is also strange that no one seems to use a Y-shaped adaptor in transposon based library prep. Is there any technical difficulties here?
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Old 04-01-2014, 06:35 AM   #2
cli
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Hi, I wonder if you have found a solution to the problem? Thanks.
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