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Old 05-09-2015, 09:05 AM   #17
pmiguel
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Location: Purdue University, West Lafayette, Indiana

Join Date: Aug 2008
Posts: 2,317
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Quote:
Originally Posted by DNATECH View Post
Hi Pmiguel,

the basic procedure looks like:
- 5 ul of library (2 nM to 3 nM including PhiX)
- add 5 ul 0,1 N NaOH
- add 5 ul Tris (200mM)
- add 35 ul Enzyme Master Mix
- load all 50 ul onto cBot
Ah, that's very interesting. They were finally forced to kick that ridiculous 50X dilution/neutralization step to the curb.

So you cluster at 200-300 pM. About 10-15x what we use on our HiSeq2500.

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Phillip
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