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Old 03-18-2014, 02:30 PM   #1
ECO
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Location: SF Bay Area, CA, USA

Join Date: Oct 2007
Posts: 1,358
Default Eliminating dry-down/speed-vac step in Nimblegen hybridization capture setup

I wanted to share with the community a small protocol modification that dramatically speeds up Nimblegen hyb setup and also makes it possible to do in much higher throughput and with lower concentration libraries.

Nimblegen's standard protocol requires that one mix together libraries, blocking oligos, and Cot-1 DNA...then evaporate it to nothingness with a speedvac. This is time consuming, messy, and annoying.

Instead of evaporating all of this, we bind it all to AMPure beads (2x cut), wash the beads once in 70% ethanol, dry them, and resuspend them in 10.5ul of hyb buffer. It takes all of 5 minutes, and no cross-contamination prone speedvac step!
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