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!
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!