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Like many folks working on low diversity libraries, we have been hit hard by the problems associated with the V3 kit - lots of time and effort wasted. Is this problem also common for high diversity libraries? I would appreciate any insights.
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I've had problems with high-diversity (gDNA) libraries with the 600-cycle V3 kits, even with a cluster density of 1K. Read 2 just tanked after about 50 bp. We are advising people not to run them at all until Illumina sorts out whatever problem this is. -
Thank you! Will it be possible to mix our MiSeq libraries with our HiSeq libraries (e.g., RNA-Seq, have not started yet) and running the mix libraries with HiSeq2500 or 3000?Comment
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I've run libraries on the MiSeq (usually for troubleshooting purposes) and then loaded them on a HiSeq once I was sure everything was okay with them. The loading concentrations are usually different, in my experience, but that should be the only issue you'll need to address.Comment
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Good to know-thanks!Comment
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Love this - good data definitely starts from good input, and poor input can only give relatively poor data. I particularly like the mention of Nanodrop/absorbance based methods for quantification. It's such a toss up if you'll get an accurate reading or what amounts to a randomly generated number, and a lot of library/sequencing related issues can be traced back to poor quant.07-01-2026, 11:43 AM
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I’m not a sequencing expert. I’m a purification scientist who uses NGS to evaluate workflows my group develops. With this perspective, we think about the sample first and the NGS workflow second. The sequencer is an exceptionally honest reporter, but it can only report on what you give it, so whether you get clean, interpretable data from an NGS workflow is largely determined before you begin.
Here are nine questions we think about, in roughly the order they matter, before...
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