Bead Recycling

[thefabnab]

Has anyone had any success working with on bead chemistries?
I'm thinking specifically doing a bead cleanup following end repair and keeping the samples on beads during adaptor ligation then resuspending those beads again in PEG to recycle the beads for another round of cleanups.
Basically, using the same beads for 2 rounds of cleanup, has anyone done this?

[atcghelix]

We always do that with our libraries and Kapa LTP library prep kits. You resuspend beads in water then add PEG at specific fractions to wash away small fragments/adapters/dimers/etc...

Essentially following the protocol here: http://protocols.readthedocs.io/proj...rep/en/latest/

[nucacidhunter]

I am surprised that on bead library protocol is still being used. There are various new efficient and less labour intensive chemistries with lower input requirement that perform much better than on bead protocols and require only one or two bead clean up after ligation.

[pmiguel]

I am surprised that on bead library protocol is still being used. There are various new efficient and less labour intensive chemistries with lower input requirement that perform much better than on bead protocols and require only one or two bead clean up after ligation.

"On bead"? Are you thinking of actual "create libraries tethered to the surface of a bead" methodologies? I didn't /think/ that was where the OP was coming from...

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Phillip