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Old 07-22-2020, 07:45 AM   #1
Anmar06
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Default Expired reagents (NEB & ligation)

I was wondering if anyone has details on how reliable expired ligation kits are? I inherited an MSc project that was dropped due to covid and most reagents in the lab are 3+ months old. I understand this is the expiry limit, as confirmed by all the stored flowcells being bone-dry, but I was wondering if this would be the case for the rest of the end-prep/ligation kits (DNA, 1D, non-PCR, no barcoding). Im brand new to this site and partially to Nanopore so I'd really appreciate all the help I can get!

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Old 07-22-2020, 01:26 PM   #2
luc
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Three months should not make a meaningful difference with these reagents - if they were stored properly.
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Old 07-24-2020, 01:26 AM   #3
Anmar06
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Quote:
Originally Posted by luc View Post
Three months should not make a meaningful difference with these reagents - if they were stored properly.
Thank you so much for your help! How about a year past expiry for the Ligation kit and about ~8 months for the NEB kit? Im already having to fight about the price of the defunct flowcells so I'd rather replace as little as I can. It's all been stored properly and unopened since first use.
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Old 07-24-2020, 11:53 AM   #4
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In our experience the ligation and associated reagents is the most sensitive part of library preps. I would expect other reagents to last longer past the expiration date, but I won't make any predictions.
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Old 07-24-2020, 12:36 PM   #5
Mel d.
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I had this discussion with nanopore tech support a couple of weeks ago. Apparently the ligation kit shelf life used to be 6 months and it was shortened to 3 months: it's the motor protein AMX (or AMII in NB kits) that is not as stable. That said, I have used kits for six months without any big problems, it's just that if they don't work you can't complain!
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Old 08-24-2020, 02:39 AM   #6
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I recently did a couple MinION of runs with a sequencing kit that was 6 days at room temp during shipment, before being stored beyond its expiration date due to the pandemic, with flow cells that were also expired. Only the NEB reagents had not expired. The runs were very good, in fact among the best we have had. It seems like a really robust system.
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Old 09-03-2020, 07:33 AM   #7
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Thanks everyone My runs have been working alright!!
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