Novel method for extracting recalcitrant plant tissue DNA
A plant tissue, stubborn technology, applied in the field of DNA extraction, can solve the problems of limited amount of DNA extraction from pear tissue, low DNA purity, cumbersome operation, etc., and achieve the effect of less extraction steps, high DNA yield and more impurities
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[0039] (1) Take 0.6 g of pear pulp tissue samples and place them in a mortar pre-cooled with liquid nitrogen, and quickly grind them to a powdery state. During the grinding process, the DNA is prevented from being degraded, and liquid nitrogen is continuously added during the grinding process. Transfer the ground powder into a 2mLEP tube, add 600μL sodium laurate extraction buffer, and shake repeatedly and slowly;
[0040] (2) Add 600 μL of a mixture of phenol, chloroform and isoamyl alcohol (25:24:1), shake repeatedly and slowly, centrifuge at 4°C, 12000 rpm for 20 minutes, and absorb 1200 μL of the supernatant;
[0041] (3) Add 840 μL of isopropanol to the supernatant obtained in step (2), slowly turn it upside down several times, let it settle for 1 hour, centrifuge at 4°C, 12000 rpm for 5 minutes, and pour off the supernatant;
[0042] (4) Wash the precipitate with 75% ethanol, gently shake and wash repeatedly, centrifuge at 10,000 rpm at 4°C for 10 minutes, discard the su...
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