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I have an undiluted rapid library that was prepared 4 months ago. Does anyone have experience with sequencing old(er) libraries?
Thanks.
Barry
Thanks.
Barry
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I don't have direct experience with older libraries but if the library was free from nucleases there is no reason why it shouldn't work. I would run it on the bioanalyser and qubit beforehand to verify that it is intact. -
We have sequenced libraries almost 1 yr old without a problem. I was having a discussion about library stability with support this past few days. There is no official recommendation about how long an undiluted library can be stored for. The 10e07 molec/ul dilution is fine to up 2 months and shouldn´t be frozen-thawed more than once. However I believe there is variability from library to library, and also everything depends on how well it has been stored.
If makes you feel safer, do like barkasn said and run an HS chip beforehand.Last edited by MissDNA; 05-20-2012, 05:56 AM.Comment
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Thanks for the input/suggestions. I'll check it out on the bioanalyzer and maybe re-measure the concentration.
BarryComment
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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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