I stored my (ear clipping) tissue samples at -20 to run a genotyping PCR in a few days and I accidentally thawed the samples by leaving them out overnight. Will the tissue even be worth running? I was considering isolating the samples and nanodropping them.
I also unfortunately left an aliquot of Proteinase K at room temp overnight. Should this be thrown away as well?
Thanks!
Hello Madison,
Not sure about your DNA. I know that readily extracted DNA is quite stable even for long periods if properly stored. My guess would be that it's worth a shot if it was just one night at room temperature.
About PK... I've been working a lot with it lately because of the pandemic and i'm pretty sure it still works after an overnight stay at room temperature. We had PK from various manufacturers and everytime there were some simple rules among which
1) do not freeze/thaw multiple times
2) use the PK aliquot in less than 24h or discard it.
I can go as far as to say that some extraction kits from Promega were delivered and stored at room temperature, yet the PK was already in solution. However, they too recommended to use it in the next 24h after uncapping.
Best way, in my opinion, is to try and see what happens. You may lose some time and materials, but this kind of lessons are more valuable than 10 written protocols and data sheets because no one's going to publish those little lab tricks that will make you be the fairy of your own lab.
Hi Madison,
I guess the tissue can still be used for genotyping by PCR, there should be still enough DNA. Also the proteinase K should be still working, maybe increase the amount you use by 50% for the next digestion.
Good luck!
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