Method for extracting total DNA (deoxyribonucleic acid) of fungal hyphae
An extraction method, fungal technology, applied in the field of molecular biology research of fungi, can solve problems such as difficult acquisition of liquid nitrogen and dry ice, inability to obtain wall removal effect, easy loss of fungal hyphae, etc., to avoid labor intensity and cross-contamination , Easy to operate, prevent cross-contamination effect
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[0046] (1) Take three 2mL sterilized centrifuge tubes and add 1.3ml of DNA extraction solution respectively;
[0047] (2) Pick about 1.0g of Fomes fomentarius (L.) J.J.Kickx, Laetiporus sulphureus var.miniatus (Jungh.) Imazeki and Agrocybe cylindracea ( DC.: Fr.) R. Maire) mycelium, put it into a centrifuge tube, and proceed as follows:
[0048] (3) Use a sterilized inoculation stick to mash the mycelia as much as possible, vortex, and mix well;
[0049] (4) Place the sample in a -80°C refrigerator for 5-10 minutes, take it out and put it in a 65°C water bath for 20 minutes, take it out and vortex it every 5 minutes during the water bath, and repeat this step for 2-3 times;
[0050] (5) Add 50 μL of proteinase K (10 mg / mL) and 50 μL of lysozyme (100 mg / ml), vortex and mix well, then place on a shaker at 37°C and 200 r / min for 30 min;
[0051] (6) Add 200 μL of 20% SDS solution, vortex and mix well, put it in a 60°C water bath for 1.5 hours, and shake once every 30 minutes du...
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