How fast does cDNA degrade at room temperature

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omgboy
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How fast does cDNA degrade at room temperature

Post by omgboy » Wed Apr 04, 2007 6:58 am

Hey i need a little help with this cause i apparently left my supervisors box of 30 or so cDNA tubes for approximately 15 hours at room temperature overnight and i need to know how badly they will be degraded. I did an RT and had stored cDNA in the PCR machine for the whole weekend before at 11 degrees ( i know its wrong). Would problably try to run an electrophoresis to see how badly degraded it is but does anyone know erm how bad the degradation might be statistically or something?

blcr11
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Post by blcr11 » Wed Apr 04, 2007 5:36 pm

I'm not sure exactly what you did. If it was just a matter of leaving out some plasmids that contain cDNA inserts, then you probably haven't done much to damage anything, provided the samples aren't contaminated with nuclease, though I don't recommend it, either. If the cDNAs were just that, cDNA, then I assume they were meant to be libraries and you were trying to clone a gene from a library of some sort. Running the library on a gel probably won't tell you much as it will be a big smear anyway. Again, since it is DNA, you probably haven't damaged much by leaving it at room temperature. I don't know how comforting that will be to your advisor.

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canalon
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Post by canalon » Wed Apr 04, 2007 7:25 pm

Depends also in what your DNA was stored, if it was in some sort of Buffer, or even better TE you are probably safe. In water, which is acidic, DNA can get degraded at RT (depurination).
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