Reagent and method capable of simply and effectively extracting plant pathogenic fungus genomic DNA
A plant pathogenic fungi and genome technology, applied in the biological field, can solve the problems of long consumption time, cumbersome process, and many impurities, and achieve the effects of good integrity and stability, simple and effective extraction, and high-efficiency extraction.
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[0035] A simple and effective method for extracting genomic DNA of plant pathogenic fungi, comprising the steps of:
[0036] (1) Get 0.2 g of Sclerotinia mulberry hypertrophic sclerotinia-Mulberry mulberry, grind it to powder with liquid nitrogen, and quickly transfer it to a sterilized centrifuge tube;
[0037] (2) Add 600 μL of solution A, vortex for 1-2 minutes, and let stand for 10 minutes; the formula of solution A is as follows: 200mM Tris-HCl (pH8.0), 250mM NaCl, 25mM EDTA, 0.5wt% SDS, 1wt% β-mercapto Ethanol, RNase 8 μL.
[0038] (3) Add 260 μL of solution B to the centrifuge tube in step (2), mix well, let stand for 10 minutes, and centrifuge at 12000 rpm for 5 minutes; the formula of solution B is: 5M KAC;
[0039] (4) Take the supernatant from the centrifuge tube in step (3) into a new centrifuge tube, add an equal volume of isopropanol, mix well, let stand for 2 minutes, and centrifuge at 12000 rpm for 5 minutes;
[0040] (5) Discard the supernatant in the centri...
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