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Old 02-14-2013, 04:56 AM   #14
pmiguel
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Location: Purdue University, West Lafayette, Indiana

Join Date: Aug 2008
Posts: 2,317
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Not really damaged, just denature. Back in the early SOLiD days, there was a lot competition for genome coverage between ABI and Illumina. Seems like one consensus arrived at was that anything that heated DNA lead to worse coverage for extremely AT-rich regions. For what it is worth... I did not actually carefully read the resultant literature, so I can't say how well this claim is supported.

Anyway, around that time, ABI added a room-temp method for dissolving agarose for minelute (quiagen). They just added extra gel dissolving buffer (chaotrope.)

Since that point, I try to avoid high temp steps in genomic DNA library preps. But realistically it is not always possible. The zymo kit, aside from heating buffer, also recommends doing multiple elutions. So maybe you can get the same effect from multiple room temp elutions as you can from single high temp elutions?

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Phillip
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