I want to identify few tiny mushrooms on 28s or 18s basis, but unable to isolate them on PDA or MEA medium, and it is difficult to bring samples to my lab. Is there any method to isolate DNA from fruit body of Mushroom? Is it possible to keep fruitbody in DNA extraction buffer for a week (at Temperature around 30-34)?
Of course, DNA extraction from fuit body is possible and can be easily done by the number of kits and other techniques. For exampe you may use Qiagen DNA Extraction Mini Kit or if you need smth more simply - CTAB method of DNA isolation (DOyle, Doyle, 1987).
Storing furuit bodies in DNA extraction buffer isn't allowed and not recommended. Better way - simply to dry the specimens or store it in special fixators which allows further DNA extratcion.
Very easy to isolate DNA directly from fruiting bodies. Collect them and immediately transfer them to liqN2. Take them to lab and crush in liqN2 and add ctab buffer, isolate dna using Murray and Thompson 1980 method.
Dear Prakash you can get some idea from following papers:
http://onlinelibrary.wiley.com/doi/10.1046/j.1472-765x.2002.01071.x/pdf
http://www.currentscience.ac.in/Volumes/104/03/0345.pdf
http://www.fgsc.net/fgn44/weiland.html
https://www.researchgate.net/publication/11487234_A_rapid_and_efficient_assay_for_extracting_DNA_from_fungi
Article A rapid and efficient assay for extracting DNA from fungi
I am getting red colour pigmentation on N2 crushing of off-white fruit body
Dear Prakash
I have the apprehension that pigmentation in the DNA preparation may create
problem in PCR amplification.Although, I haven't worked on mushrooms, but this problem arises with other ascomycetous and mucoralean fungi where pigment creates a lot of problem in many downstream applications. I think extracting DNA from young mushroms can help you,otherwise, if concentration isnot such a big issue in your case, you can dilute your DNA enough so that pigment conc. gets reduced to the level which may not pose any problem to you.
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