Whenever I do DNA extraction from Ascochyta, at a stage when I want to dissolve the DNA in water, the DNA is partially or do not dissolve at all. Anyone may help me?
Use TE buffer instead of water hope you find good. TE buffer will also increase the stablity of your DNA
@ michael, i will try ur suggestions might help.
@ Irfan, what you are saying is at the last step of DNA isolation, where TE buffer is used and stored as such. but my problems starts before that. after precipitating alcohol washing, and the dissolving with pure water. Here my DNA doesnt dissolve and face difficulty in subsequent RNAse treatment.
Too much drying of the pelleted DNA in vaccum drier or at 37 C will land you into the current situation. Reduce the drying period and check if the same problem continues
if you precipitate with ethanol, dry it until you a see a pellet that looks a little more viscous than a water drop...starting to get cloudy. it's a fine balance but for ethanol precipitation, you don't need to wait until it is a white precipitate (or at least I didn't). I usually add water JUST before that happens and it works well. Alternatively, heating will help (but personally, I was afraid on precipitate that was too small to see remaining out of solution since the starting pellets are already so tiny.)
If you over dry the DNA it will be difficult to dissolve. What you can do add DDW and incubate at 60 degree Celsius for 30'-45' and hope fully it will be dissolved.
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