hello .
I have a lysis solution containing RNaseA for dna extractoin.
I have stored the solution in a refrigerator for a long time.
For that reason, the solution now shows some white pellets.
How can I resolve the white pellets???
Hello Jong-Hwan Shin,
The best would be to make a fresh new lysis solution.
Another question: does this lysis solution contains NaOH for alkaline lysis of bacterial cells ? If yes, and that it was stored in glass and not in plasticware, this white pellet could be sodium silicate.
One fast way to proceed would be to cenrrtifuge the lysis solution and test it for activity! You coudl be lucky and have a still functional lysis solution after all.
RNaseA is a very very stable enzyme so ...
Best wishes and good luck for your experiments
Pr Philippe Urban
Dear Jong, I am not sure if your lysis solution contain SDS? In that case it will be good idea to warm up your solution by 45-50C on a watter bath to see if you can redissolve the SDS precipitates. Generally SDS tends to fall out of the solution at low temperature. So you may try your luck by warming the solution. I agree with Filippe that RNaseA is extremely stable enzyme, so warming up should not pose a problem. Secondly if your solution contain NaOH in a glass bottle, than as Filippe suggested, it could be the crystals of Sodium silicate. Then the better option will be either to centrifuge it and use supernatant for the lysis OR else go with the fresh lysis buffer and discard the old one. Good luck with your experiments.
Hi Jong
I agree with Rajesh, most lysis buffers contain SDS which precipitates when cold. This should be a problem provided you bring to no higher than 50 C. I advise room temperature thawing. It takes longer but it's safer really. Once the precipitate has dissolved you can carry on as per protocol.
As both Rajesh and Phillipe mentioned, the NaOH may also be a problem. This, however, depends on the concentration used which depends on the protocol being followed.
Just as a precaution - I never use a lysis buffer older than 2 weeks (when stored at 4 C in interim).
Regards,
Nathaniel
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