Hey,
In order to standardize and optimize positive controls for Real-time PCR in our laboratory, and also to gain time, we were wondering the stability of plasmid dilutions for semi to long term use in our diagnosis routine.
We want to know if we can store our positive controls dilutions at 4°C for a month without loosing any stability ?
I'll appreciate any knowledge or experience or feedback on that subject
cheers.
What I've usually done is made aliquots of my plasmid DNA in strip tubes and stored them at -80*C. Pull out a set of tubes, thaw, use and then toss any remaining. Don't do multiple freezing and thawing cycles. It's worth the bother of aliquoting so you know you have consistency
Hi,
Extending a little on both answers, another determinant of course is the purity, mostly deproteination, of your DNA. CsCl-density-gradient-purified DNA will be stable for months to years at 4°C, the DNA of minipreps may become degraded already overnight.
Best wishes, Ernst
Thanks for the kind answers,
I asked for a little bit more precision about purification of plasmids from the company that does this job for our laboratory.
They told me the plasmids were purified with the kit NucleoSpin Plasmid, Macherey Nagel, Ref. 740588250 in accords with the instructions of the kit.
I guess that means that they are minipreps and should be story at -20°C for long time storage (1-2months) ? Any feedback or experience would be really appreciated.
thank you in advance for your answers.
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