Method for effectively extracting genomic DNA (deoxyribonucleic acid) of melaleuca alternifolia
A Melaleuca alternifolia and genome technology, applied in the field of plant DNA extraction, can solve the problems of low DNA yield, poor DNA quality, and high cost, and achieve the effects of enhancing safety, saving costs, and saving experimental operation time
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[0033] Example 1:
[0034] A method for effectively extracting the genomic DNA of Melaleuca alternifolia, specifically comprising the following steps:
[0035] (1) Grind 1 g of fresh leaf samples of Melaleuca alternifolia with liquid nitrogen to powder, quickly transfer it to a centrifuge tube, add 600 µl of conventional pre-warmed DNA extraction buffer, mix well and place in a water bath at 65°C for 30 minutes, in which DNA extraction buffer The formula of the solution is: 2% CTAB, 100 mM Tris-HCl with pH 8.0, 20 mM EDTA with pH 8.0, 1 M NaCl;
[0036] (2) Take out the centrifuge tube and cool it to room temperature, add the mixture of chloroform and isoamyl alcohol with the same volume as DNA extraction buffer, the volume ratio of the mixture is: chloroform: isoamyl alcohol = 24:1, and after mixing, add it at 12000 r / min centrifugation for 10min;
[0037] (3) Take the supernatant and place it in a new centrifuge tube, add the same volume of chloroform as the supernatant, ...
Example Embodiment
[0043] Example 2:
[0044] A method for effectively extracting the genomic DNA of Melaleuca alternifolia, specifically comprising the following steps:
[0045] (1) Grind 2g of fresh leaf samples of Melaleuca alternifolia with liquid nitrogen to powder, quickly transfer it to a centrifuge tube, add 800µl of conventional pre-warmed DNA extraction buffer, mix well and place in a water bath at 65°C for 30 minutes; the DNA extraction buffer The formula of the solution is: 2% CTAB, 100 mM Tris-HCl with pH 8.0, 20 mM EDTA with pH 8.0, 1 M NaCl;
[0046] (2) Take out the centrifuge tube and cool it to room temperature, add the same volume of chloroform and isoamyl alcohol mixture as DNA extraction buffer. / min centrifugation for 10min;
[0047] (3) Take the supernatant and place it in a new centrifuge tube, add the same volume of chloroform as the supernatant, mix well and centrifuge at 12000 r / min for 10 min;
[0048] (4) Take the supernatant and place it in a new centrifuge tube, a...
Example Embodiment
[0053] Example 3:
[0054] A method for effectively extracting the genomic DNA of Melaleuca alternifolia, specifically comprising the following steps:
[0055] (1) Grind 1.5 g of fresh leaf samples of Melaleuca alternifolia with liquid nitrogen to powder, quickly transfer to a centrifuge tube, add 700 µl of conventional pre-warmed DNA extraction buffer, mix well, and place in a water bath at 65°C for 30 minutes; DNA extraction The formula of the buffer is: 2% CTAB, 100 mM Tris-HCl at pH 8.0, 20 mM EDTA at pH 8.0, 1 M NaCl;
[0056] (2) Take out the centrifuge tube and cool it to room temperature, add the same volume of chloroform and isoamyl alcohol mixture as DNA extraction buffer. / min centrifugation for 10min;
[0057] (3) Take the supernatant and place it in a new centrifuge tube, add the same volume of chloroform as the supernatant, mix well and centrifuge at 12000 r / min for 10 min;
[0058] (4) Take the supernatant and place it in a new centrifuge tube, add 1.5 times ...
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