Usually we keep extract overnight after adding 1/10th volume of sodium acetate and 2 volumes of ethanol. My question is, if we keep it at -20 Celsius for 3 days, will it affect DNA quality? We are yet to do ethanol wash.
The sodiumacetate neutralizes the ionic charge of the phosphate backbone of the DNA and also builds helps destroying the hydrate hull around the DNA (the acetate anion needs water to stay in solution). Together with the ethanol this precipitates the DNA.
No problem at all. Actually you can keep DNA in 70% Ethaool for a long time as your DNA will be in the solid phase and no enzymatic reaction can affect it.
You can. Or you can finish the precipitation and freeze it in TE buffer (or water).
Ethanol is a precipitating agent and NaAc is protecting agent from enzymes.
I would recommend -70 rather than -20. I have done this many times for more than a week with no problems
The sodiumacetate neutralizes the ionic charge of the phosphate backbone of the DNA and also builds helps destroying the hydrate hull around the DNA (the acetate anion needs water to stay in solution). Together with the ethanol this precipitates the DNA.
Before going to use for experiment, it's better to conform the required ph
Sometimes, I stored that solution for 1h, sometimes for overnight. They are OK as well. I never store them for 3 days, but I think that there's no problem.
BR
i usually precipitate DNA by incubating at -80C for one hour and it worked well for me, In the sam e time , i tried to incubate overnight at -20C and i didnt have any problems in both the kind of precipitation.
I usually do 30 min at -80, and then another 30 min at -20 when I want to get the procedure done. Works every time. But storing in -20 for 3 days is not a problem. You'll get lots of DNA.
It is ok to keep it for 3 days, We usually keep it in -20 overnight when we dont get the spool of DNA when chilled ethanol is added. Can you still see the spool of DNA? if yes the just trasnfer it to an a fresh eppendorf tube and go for ethanol wash. if you cannot see it centrifuge it and you will find the DNA on inner side of the wall of the centrifuge tube.
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