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-   -   Problem with new Hi-Q Chef kit for proton (http://seqanswers.com/forums/showthread.php?t=61465)

ptery 07-23-2015 08:15 AM

Problem with new Hi-Q Chef kit for proton
 
With the discontinue of the old PI IC kit, we have to switch to new Hi-Q chef kit. We had great success with the old PI IC kit, but facing dramatic drop in loading percentage and read quality with the new kit. Any one have experience with new Hi-Q chef kit?

NextGenSeq 07-23-2015 08:58 AM

We had a problem with an NEBNext Ion Torrent library but the Hi-Q worked fine with a library made using Ion Torrent reagents.

What library concentration are you using? We use 85 pM.

ptery 07-23-2015 09:59 AM

I use 70ul @ 100pM for the old IC kit. For Hi-Q kit, I first followed the protocol with 25ul @ 50pM with only 49% loading. So I increased loading to 25ul @ 200pM, the result is still not good. Finally I tried 25ul @ 400pM, none of those conditions give me good yield. (45M/chip at the most for 400pM loading).


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