A simplified extraction method of total dna from blueberry fruit
An extraction method and fruit technology, applied in the field of DNA extraction, can solve the problems of affecting the purity, cumbersome operation, not taking into account the differences of different samples, etc., and achieve the effects of avoiding pollution, high safety, and time-consuming
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Embodiment 1
[0022] A simplified blueberry fruit total DNA extraction method comprises the following steps:
[0023] S1, Material pretreatment: Collect 1g of blueberry fruit, mash it, add 5ml of 0.1mol / l sodium hydroxide solution, grind it into a slurry, incubate at 65±1°C for 10min, shake 3 times during the period, add 0.1mol / l formic acid The pH of the solution is adjusted to 6.0 to obtain pretreated blueberry pulp;
[0024] S2, add 1ml of the mixed solution to 1ml of the pretreated blueberry pulp, and the ratio of 0.1mol / l formaldehyde solution, vitamin C and DNA extract in the mixed solution is 1ml:0.01g:20ml, water bath at 37±1°C for 20min ;Centrifuge at 12000rpm for 10min, collect the supernatant;
[0025] Among them, 1L DNA extract contains 0.1mol Tris-HCl, 0.1mol EDTA, 1.5mol NaCl, 10gCTAB, the solvent is double distilled water, pH 8.0;
[0026] S3, add isopropanol equivalent to 0.6 times the volume of the supernatant that has been pre-cooled at 4°C to the supernatant, mix well, ...
Embodiment 2
[0029] A simplified blueberry fruit total DNA extraction method comprises the following steps:
[0030] S1, material pretreatment: collect 2g blueberry fruit, crush it, add 10ml 0.1mol / l sodium hydroxide solution, grind it into a slurry, incubate at 65±1°C for 10min, shake 4 times during the period, add 0.1mol / l acetic acid The pH of the solution is adjusted to 6.0 to obtain pretreated blueberry pulp;
[0031] S2, add 2ml of mixed solution to 2ml of pretreated blueberry pulp, and the ratio of 0.1mol / l formaldehyde solution, vitamin C and DNA extract in the mixed solution is 1ml:0.05g:30ml, water bath at 37±1°C for 20min ;Centrifuge at 12000rpm for 10min, collect the supernatant;
[0032] Among them, 1L DNA extract contains 0.1mol Tris-HCl, 0.1mol EDTA, 1.5mol NaCl, 10gCTAB, the solvent is double distilled water, pH 8.0;
[0033] S3, add isopropanol equivalent to 0.8 times the volume of the supernatant that has been pre-cooled at 4°C to the supernatant, mix well, place at -20...
Embodiment 3
[0036] A simplified blueberry fruit total DNA extraction method comprises the following steps:
[0037] S1, material pretreatment: collect 2g blueberry fruit, crush it, add 10ml 0.1mol / l sodium hydroxide solution, grind it into a slurry, incubate at 65±1°C for 10min, shake 4 times during the period, add 0.1mol / l acetic acid The pH of the solution is adjusted to 6.0 to obtain pretreated blueberry pulp;
[0038] S2, add 2ml of mixed solution to 2ml of pretreated blueberry pulp, and the ratio of 0.1mol / l formaldehyde solution, vitamin C and DNA extract in the mixed solution is 1ml:0.03g:25ml, water bath at 37±1°C for 20min ;Centrifuge at 12000rpm for 10min, collect the supernatant;
[0039] Among them, 1L DNA extract contains 0.1mol Tris-HCl, 0.1mol EDTA, 1.5mol NaCl, 10gCTAB, the solvent is double distilled water, pH 8.0;
[0040] S3, add 0.7 times the volume of isopropanol pre-cooled at 4°C to the supernatant collected in S2, mix well, place at -20°C for 45 minutes, centrifug...
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