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-   -   Eliminating dry-down/speed-vac step in Nimblegen hybridization capture setup (http://seqanswers.com/forums/showthread.php?t=41736)

ECO 03-18-2014 02:30 PM

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!

ZWB 03-19-2014 07:47 AM

ECO,

Thanks for the great idea! This will save me a lot of headache as I prepare for some higher throughput experiments I'm planning.

Zach

ECO 03-25-2014 12:49 PM

Let me know if it works for you. I hate speedvacs! :D

biaoluo 03-29-2014 07:25 PM

ECO,

Thank for the idea. Could you please clarify whether your hybridization includes the Ampure beads ?

Thanks,
BL


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