Will anybody please let me know ways of storing DNA
I'm extracting DNA from animal tissues using a kit called DNAZole and a conventionala method using lyses buffers with protienase K
The kit results in producing a pellate and is stored in TE buffer
it reccamends H2O or 8mM NaOH
What ever did that pellet doesnt get dissolved
Will any body please help me with your experiences
Thanks
dilip
I usually dissolve DNA in TE buffer and store at 20C or 4C depending on my usage. You want to avoid excessive freeze-thaw of the sample.
Reference I just came across discussing TE vs water for storage of DNA.
http://www.dnagenotek.com/pdf_files/PDPR012_LongTermStorage.pdf
I usually dissolve DNA in TE buffer and store at 20C or 4C depending on my usage. You want to avoid excessive freeze-thaw of the sample.
Reference I just came across discussing TE vs water for storage of DNA.
http://www.dnagenotek.com/pdf_files/PDPR012_LongTermStorage.pdf
My tips would be:
Once you've pelleted the DNA and washed with 70% ethanol, let it air dry - but only just long enough that the pellet turns from the easily visible white to transparent. Pellets that have airdried for too long tend to be more difficult to resuspend in TE or water.
I used to use TE all the time but after talking to a guy from PlasmidFactory I switched to using millipore filtered deionised water. His argument was that the buffer strength of the TE did not efficiently protect the DNA from degradation over time. And clearly there are fewer problems when using the DNA for downstream processing techniques that do not like the presence of EDTA (sequencing etc). But you can probably avoid this problem by keeping your DNA stocks quite concentrated and then diluting them in water for sequencing etc.
I've never heard of disolving DNA in NaOH (even low concentrations). If you have to have it in NaOH, I would resuspend it initially in water then add the NaOH. I seem to remember doing Southern blots in NaOH because you want the DNA to transfer as single-stranded. Do you need your DNA ss?
if that is genomic DNA that you are extracting, you should NOT freeze it, but instead store it at 4°C
Hi,
You may air-dry the pallet for too long and dna amount you got could be high.
Dissolving in nuclease-free water if u intend to use these samples for sequencing. Dna in water only good for short term storage. TE stablizes DNA for long term storage.
Dav