In order to carry out my genomic project, I need isolate DNA from almost 1,000 megagametophytes in a conifer species. So, probably I have to take into account a method which allow me store the gametophyte tissue for long periods. I would be grateful if anybody could drive me about good practices in megagametophyte DNA isolation.
Thank you in advance.
Hi Gustavo,
We use DNA/RNA shield by Zymo Research and it works well with adults mosquitoes, larvae, eggs... They say that it can preserve any type of DNA, including plant tissue or cells.
We store it at RT for weeks and at -4 or -20 for months, never saw any DNA degradation.
https://lucerna-chem.ch/media/downloads/Zymo/DNA-RNA%20Shield-Brochure.pdf
you can store at -80 for a months or as experience of Umberto store it at -20.
I preferred storage condition -80.
In my opinion, its better to freeze dry the samples so that you can keep them at normal freezer. If you do not have this facility, then you can store the samples at - 80C for long time without adverse effect on DNA. I have stored samples (leaf and seed) at -80C for more than 6 months and isolated DNA and performed PCR without any problem.
Thanks so much for all your advices! After some essays, I concluded that in the case of conifers megagametophyte, It is pretty recommendable do not store the tissue for long time before DNA isolation. Actually, I get excellent results when I isolated the DNA at the same day (!) that I got the tissue. Regarding the DNA isolation protocol (CTAB2x or DNeasy plant mini kit 250), I consider there were not so much variation.
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