Does anyone have experience sequencing small RNA libraries on the NextSeq? We are consistently getting low cluster densities (140-150K/mm2) even with a 10% PhiX spike-in to increase diversity. The libraries were made with the NEBNext Small RNA Kit.
There is clearly low diversity in the libraries by looking @ the post-run specs, so I thought that increasing the PhiX while leaving loading concentration the same would help the issue (Quality scores have been great with %Q30 above 80%). What I actually found was that increasing the PhiX any more than 10% dropped cluster density even further.
I would truly appreciate any thoughts on this. Thanks!
There is clearly low diversity in the libraries by looking @ the post-run specs, so I thought that increasing the PhiX while leaving loading concentration the same would help the issue (Quality scores have been great with %Q30 above 80%). What I actually found was that increasing the PhiX any more than 10% dropped cluster density even further.
I would truly appreciate any thoughts on this. Thanks!
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