Hello and Good day everyone.
Previously, I did DNA extraction from gram positive bacteria by boiling method. After that, I preserved it at temperature -20 degree which is very suitable for DNA preservation. But, I has bit doubt whether that DNA will degrade due to contamination since I used boiling method. Has anyone experience to deal with this situation? Thank you.
Dear Abdul Rahim,
excessive boiling can only denature DNA. Normally for DNA isolation we heat from 60-70 degree Celsius where it may not degrade the DNA.
I have never preserved DNA extracted by boiling method. I usually used it for 1-2 PCR.
It depends what you are going to do with it. I've kept boil extracted DNA form bacteria for weeks or months at 4oC and the PCR results have been just fine. I actually find that in general DNA goes off more at -20oc and keeps better at 4oc or -70-80oC.
It is better if you keep it at -20 or -80ºC (for long term. years). But I recommend you avoid multiple freezing, for example doing aliquotes.
Hi! I agree with Hector. If you are going to use the DNA sample several times, just save it into the fridge or doing aliquotes. I have many samples of DNA that I have obtained from gram positive bacteria also. I did them on 2012 and last year I used them for a PCR and everithing worked well.
good luck
Graciana
I strongly agree with Hector. I would also avoid boiling.Your best off storing your DNA samples at -20 or -80ºC.
Better not to preserve, each time use fresh. otherwise non-specific bands will be there.
Several times I used in PCR drosophile genome DNA after 5-6 month in a freezer at -20C. PCR reaction worked fine.
Unfortunately, I don't know how stable is DNA after boiling.
A protocol on DNA extraction by boiling method includes freezing at -20 for 10 minutes. what effect does it have on the quality of DNA or yield in case one does not freeze and thaw?
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