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Old 02-12-2013, 06:58 AM   #8
pmiguel
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Location: Purdue University, West Lafayette, Indiana

Join Date: Aug 2008
Posts: 2,317
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Quote:
Originally Posted by razibus View Post
Thank you for your answer, it's very relevant.

I tried Qiagen columns but also Zymo Research columns. Both works perfectly on PCR amplicons but poorly on sonicated gDNA.

I will give a try to Ampure XP. Nevertheless, the elution volume is ~30 and 40Ál, and I need only 10Ál. (I don't like SpeedVac very much)

Anyway, I don't get why we lost so much gDNA in purification columns...
Once you get above a certain size, some clean up columns hold more tightly onto DNA and hence will not elute the DNA. We were previously unable to find any clean up column that would "let go" of DNA larger than 9 kb or so. This seems to be a characteristic of silica based methodologies.

Does Ampure have this issue? Not sure we have tested it, but probably not. Ampure gently binds DNA onto its magnetic beads via carboxyl groups in the presence of a precipitant (PEG).

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Phillip
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