High-efficiency extraction method of peanut leaf DNA
An extraction method and leaf technology, applied in the field of high-efficiency extraction of peanut leaf DNA, can solve the problems of high extraction cost, low extraction efficiency, and low cost, and achieve the effect of good DNA extraction quality, high extraction efficiency, and simple operation
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[0022] A method for efficiently extracting peanut leaf DNA, comprising the steps of:
[0023] (1) Collection of peanut leaf samples: take about 0.2 g of fresh young leaves, and quickly put them into a 2ml centrifuge tube for cryopreservation;
[0024] (2) Grinding of peanut leaves: at room temperature, add 300ul 2wt% CTAB solution (add 1wt% β-mercaptoethanol before use) to a centrifuge tube, add steel balls, use a tissue grinder 60HZ to grind the sample for 60s (60s can grind the leaves Grind it fully while avoiding sample oxidation to the greatest extent), and then add 600ul CTAB solution (add 1wt% β-mercaptoethanol before use), and put it in a -20°C refrigerator for long-term storage;
[0025] (3) Water bath: put the sample solution obtained in step (2) into 65°C water for 30 minutes, and shake the sample solution once every 5-10 minutes;
[0026] (4) Use chloroform / isoamyl alcohol (volume ratio 24:1) solution for the first extraction: add an equal volume (900ul) of chlorof...
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