Blue pippin size selection elution volume

[anna_m]

Hello,
Has anyone has tried eluting their TruSeq DNA libraries in a smaller volume than the 40uL stated in the protocol?

We carry out size selection on our TruSeq nano libraries after the PCR enrichment step which works for us but gives very low concentrations. I was wondering if we eluted in perhaps half the volume, we might get higher concentrations for running on the sequencers.

It would mean that the test step prior to beginning a run would technically be a fail as it tests volumes in both the loading and elution wells. Do you think that would be a big issue?

Thanks in advance for your thoughts!

[nucacidhunter]

There are two options:

1- SpeedyVac concentrate Pippin eluate
2- Column or bead clean and elute in lower volume

[kmcarr]

Are you using a 2% cassette (100-600bp) to size select your libraries? Sage sells a version of the 2% cassette which has a 25ul elution volume instead of the standard 40ul volume. Product# BSF2010 (http://www.sagescience.com/wp-conten...6_00-CD101.pdf)

[anna_m]

That's really interesting kmcarr. No, we use the 1.5% cassette because we size-select the 550bp nano libraries at 640-700bp so the 2% range would be too small.

Thanks to you and and nucacidhunter for your replies.