A high-efficiency extraction method of guava leaf genome dna
An extraction method and guava technology, applied in the field of molecular biology, can solve the problems of difficult to mention high-quality genomic DNA, can not be applied well, complex species, etc., improve the extraction rate and purity, and is conducive to popularization and utilization , a wide range of effects
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Embodiment 1
[0030] A high-efficiency extraction method of guava leaf genome DNA, comprising the following steps:
[0031] (1) Put 0.5 g of chopped guava leaves into a sterile mortar, add liquid nitrogen, use a high-frequency vibrating ultrafine powder machine to grind the guava leaves into powder, then transfer the powder into a centrifuge tube, add 1.2mL of washing liquid, after mixing, ultrasonic treatment, let stand in ice for 15min, centrifuge at 10000rpm for 10min to obtain supernatant a and bottom residue a, wherein, the composition of washing liquid includes: 50mM Tris-HCl, 5mM EDTA, 350mM sorbitol, The volume fraction is 2% PVP, the volume fraction is 0.5% β-mercaptoethanol, and the pH value of the control lotion is 8.0;
[0032] (2) Add 50 μL of cellulase and pectinase to the bottom residue a, and after standing in ice for 10 minutes, centrifuge at 10,000 rpm / min for 10 minutes to obtain supernatant b and bottom residue b;
[0033](3) Add CTAB extraction buffer to the bottom res...
Embodiment 2
[0040] A high-efficiency extraction method of guava leaf genome DNA, comprising the following steps:
[0041] (1) Put 0.5 g of chopped guava leaves into a sterile mortar, add liquid nitrogen, use a high-frequency vibrating ultrafine powder machine to grind the guava leaves into powder, then transfer the powder into a centrifuge tube, add 1.0mL of washing liquid, after mixing, ultrasonic treatment, let stand in ice for 15min, centrifuge at 10000rpm for 10min to obtain supernatant a and bottom residue a, wherein, the composition of washing liquid includes: 50mM Tris-HCl, 5mM EDTA, 350mM sorbitol, The volume fraction is 2% PVP, the volume fraction is 0.5% β-mercaptoethanol, and the pH value of the control lotion is 8.0;
[0042] (2) Add 55 μL of cellulase and pectinase to the bottom residue a, and after standing in ice for 10 minutes, centrifuge at 10,000 rpm / min for 10 minutes to obtain supernatant b and bottom residue b;
[0043] (3) Add CTAB extraction buffer to the bottom re...
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