Exonuclease I

[airun78]

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!

[Olaf Blue]

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.