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Old 04-17-2012, 12:28 AM   #1
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Location: Germany

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Default EDTA interferes with library prep?

Hi there,
we're currently preparing RNA samples for small RNA Seq on the Illumina HiSeq using the new sample prep kit from NEB. For DNA digestion, usually EDTA is used to stop the DNase reaction. Do you know if that EDTA (at a conc of 8 mM) interferes with the library prep (adapter ligation, PCRs, etc.)?
Many thanks in advance!
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Old 04-18-2012, 08:50 AM   #2
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Location: Purdue University, West Lafayette, Indiana

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I don't have details, but any reaction requiring a divalent cation will fail because quite a bit of the cation will be chelated (and thus probably not available to the enzyme). But most clean up methods will remove the EDTA, so it depends on where you plan to enter a library prep protocol.

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