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I ran an RNA 6000 nano chip with RNA samples in all 12 wells, some randomly came back flagged with error: unexpected signal in fast region. Is there any way to troubleshoot this after the fact to get an accurate RIN number?
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You should be able to manually adjust the fast region signal allowance under the advanced settings and this will clear the error, allowing for a RIN to be called. -
Agilent tech support is really helpful with fixing these problems, if you are unsure how to do so. (Via phone or email them the run file)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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