Has any one extracted DNA from the walnut leaf samples preserved in 70% ethanol? I am worried about the quality of DNA. Can it be used for further analysis like linkage mapping and studying of genetic diversity ?
It doesn't affect the DNA, after DNA wash with ethanol, discard ethanol, again centrifuge for a minute (no ethanol added), followed by drying the pellet in oven, resuspend the pellet with ddH2O or buffer.(incubate @ 65'C if needed) and store @ -20
Good luck
No. As per my knowledge/experience there should not be any problem storing samples either in Alcohol (Ethanol) or 70% alcohol (Ethanol) to extract DNA from that (I used to isolates many (hundreds). Absolutely no problem. Only thing is that you should take care to properly air dry till the total Alcohol/Ethanol is removed or evaporated. Leave the sample on tissue paper either o/n or longer times or thoroughly by using by tissue paper.
I think it doesn't affect the quality and quantity of DNA. However you should be very careful while air drying the pellets of DNA. Ethanol should be completely evaporated before resuspend in TE buffer or ddH2O.
Ethanol 70% doesn't harm DNA, but does not inhibit DNAse in raw material completely. It is also not the preservative of choice for raw plant material.
Nevertheless, for pcr it can still be fine to use.
Check Nagy 2010 for starting points: https://www.researchgate.net/publication/224944842_A_hands-on_overview_of_tissue_preservation_methods_or_molecular_genetic_analyses
and eg Saldago 2007 for genomic DNA degradation in 70% EtOH, albeit in sponges:
http://www2.ib.usp.br/~mcust/7thISS/09-book/pdf/Salgado%20et%20al%20-%20Field%20preservation%20and%20optimization%20of%20a%20DNA%20extraction%20method.pdf
HTH, René
Article A hands-on overview of tissue preservation methods or molecu...
Dear Rene van Wezel,
Thanks a lot for the information and articles. I would also like to thank Bhaskar Gouda, Jetty Ramadevi, and Shreeti Pradhan for your valuable comments
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