Method for extracting high-quality DNA (deoxyribonucleic acid) in dry leaves of soybeans suitable for transgenic detection
An extraction method and technology of dried leaves, which are applied in the field of life sciences, can solve the problems that DNA contains a lot of salt ions and impurities, cannot remove pigment substances, and cannot effectively remove polysaccharides, etc., to achieve simple extraction, obvious specific products, and transparent color Effect
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[0031] Example 1: Extraction of total DNA from dried soybean leaves
[0032] (1) The steps of the method of the present invention are as follows:
[0033] ①Take about 0.04g of dried soybean leaves, put them in a 1.5mL centrifuge tube, add 200μL of Buffer I (0.2mol / Ltris-HCl, 0.05mol / L EDTA, 1mol / L NaCl, pH8.0), soak for 10 minutes , Heated in a water bath at 60℃, cut the macerated leaves with small scissors to obtain about 0.04g of foam;
[0034] ②Add 1.2mL Buffer II (the mass ratio of sterile water to β-mercaptoethanol is 40:1), mix upside down for 5 minutes, centrifuge at 10000r / min for 5 minutes, discard the supernatant, and repeat this step 2 to 3 times. Until the supernatant is clear;
[0035] ③Take the precipitate, add 600μL of 2×CTAB extract (containing 2% by mass PVP, 1.4mol / L NaCl), 3% by mass β-mercaptoethanol, 2μL of proteinase K, shake well, water bath at 65℃ for 60 minutes, each Shake upside down every 10 minutes to get the mixture;
[0036] ④Add the same volume of pheno...
Example Embodiment
[0049] Example 2: Extraction of total DNA from dried soybean leaves
[0050] ①Take about 0.04g of dried soybean leaves, put them into a 1.5mL centrifuge tube, add 200μL of Buffer I (0.2mol / Ltris-HCl, 0.05mol / L EDTA, 1mol / L NaCl, pH8.0), soak for 15 minutes , Heated in a water bath at 40℃, cut the macerated leaves with small scissors;
[0051] ②Add 1.2mL Buffer II (the mass ratio of sterile water to β-mercaptoethanol is 40:1), mix upside down for 5 minutes, centrifuge at 10000r / min for 5 minutes, discard the supernatant, and repeat this step 2 to 3 times. Until the supernatant is clear;
[0052] ③Take the precipitate, add 600μL of 2×CTAB extract (containing 2% by mass PVP, 1.4mol / L NaCl), 3% by mass β-mercaptoethanol, 2μL of proteinase K, shake well, water bath at 65℃ for 40 minutes, Shake upside down every 10 minutes;
[0053] ④Add an equal volume of phenol:chloroform:isoamyl alcohol (volume ratio 25:24:1) mixture, mix upside down for 10 minutes, and centrifuge at 12000r / min for 10 ...
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