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Old 11-14-2012, 07:46 PM   #1
new2seq
Junior Member
 
Location: USA

Join Date: Nov 2012
Posts: 2
Default turbo dnase ribo-zero

Hello everyone
I am planning on using Turbo DNase from Ambion to treat my Trizol extracted RNA, use the DNase treated RNA for a ribo-zero kit to deplete the rRNA, and prepare ScriptSeq libraries. I am a little confused with the cleaning steps and where to do them regarding to the DNase treatment and following ribo-zero kit.
Here are my questions:
1-should I remove the Turbo DNase using a column instead of the inactivation gel (is this the consensus?), proceed to the ribo-zero kit and precipitate the RNA using Ethanol precipitation or is it better to use a column based/ the Agencourt RNA beads.

2-or should I use the inactivation gel, proceed to the ribo-zero kit and then purify the RNA using a column or Agencourt RNA beads.

Any recommendation on which workflow to use?
Thanks!

Last edited by new2seq; 11-14-2012 at 07:48 PM.
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