i would like to know that i have isolated about 30 different kind of isolates from fruit peel at harvesting stage..and have pure colonies and i want to do Identification, so i have finished microscopy but for verification want to do molecular study. so
1. how to prepare sample for fungal isolate for DNA extraction （please explain from new culture grow to the end）?
2. Best method for unknown fungal isolates DNA extraction?
3. how can store fungal isolates for about 6 to 12 months =?
I shall be thankful to you
looking forward to hearing from you.
Regard. RIZWAN.HM
1) confirm that you have pure single -spore isolates without other fungi or bacterial contamination.
2) make several replicas for long-term storage (simplest way - to keep them on sterile grain (barley, wheat) in glass tubes with cork srew at room temperature. Sterilize wet grain 2 times at 120oC.
3) use fresh, fast-growing culture, liquid culture is better, but it can be easy contaminated.
4) it is better to use liquid Nitrogen or lyophilized mycelium to destroy cell walls.
5) use 2-3 different extraction protocols if you have different species of fungi and you do not know the best method for all of them.
Dear Hafiz
This article can help you.
Article A simple method for a mini-preparation of fungal DNA
Read our paper on it Article Morphological, Pathological and Molecular Variability in Col...
https://onlinelibrary.wiley.com/doi/abs/10.1111/j.1439-0434.2005.00959.x
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