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Fully exonuclease-resistant custom oligonucleotides? memeri Sample Prep / Library Generation 5 10-31-2011 07:43 AM

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Old 07-17-2014, 08:52 AM   #1
airun78
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Location: london

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Default Exonuclease I

Hi,

I was wondering what do you think of using exonuclease I to get rid of adapter contamination from an RNA-seq library?
I am using ScriptSeq to make my libraries and although I used AMPure (1x ratio) to purify the library I still had a lot of adapter contamination. I did a 2nd purification (0.9x) which helped but when preparing another library from highly fragmented RNA I used Exonuclease to remove excess PCR primers plus MinElute PCR purification to purify the library as recommended by the kit and it has worked wonders with adapter contamination!
I was thinking of using Exonuclease I plus AMPure with all my samples.
Any thoughts?

Thanks!
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Old 07-18-2014, 09:29 AM   #2
Olaf Blue
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That should work. Epicentre includes Exonuclease I in their kit but it's only referred to for use in removing excess PCR primers in FFPE applications. I was told it was holdover from the previous version of the kit.
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