Method for quickly extracting total ribonucleic acid (RNA) from Rubus
A technology of Rubus genus and an extraction method, applied in the field of plant molecular biology, can solve problems such as long time consumption, DNA pollution, RNA loss, etc., and achieve the effects of short time consumption, increased applicability, and simplified operation process
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Embodiment 1
[0038]1) Take a 2ml centrifuge tube, add 1000 μl of sugar-free buffer to the centrifuge tube, add 5 μl of β-mercaptoethanol, and pre-cool at -20°C for 10 minutes; 0.15g, add 0.1g cross-linked polyvinylpyrrolidone, grind into powder under liquid nitrogen freezing conditions, quickly transfer to the centrifuge tube in step 1), mix gently up and down, and let stand on ice for 10min; 3) Centrifuge at 3200×g for 10 min at 4°C; 4) Completely remove the supernatant obtained in step 3), add 700 μl of cell lysate preheated at 65°C to the pellet, vortex for 1 min, and incubate at 60°C for 30 min; 5) After the mixture in step 4) is cooled to room temperature, add 700 μl of water-saturated phenol, 13 μl of glacial acetic acid, and 140 μl of chloroform in sequence, and shake vigorously for 2 minutes; 6) centrifuge at 16,000×g for 10 minutes at 4°C; ) and transfer 500 μl of the supernatant obtained by centrifugation into another centrifuge tube, add 500 μl of -20°C pre-cooled isopropanol to...
Embodiment 2
[0041] 1) Take a 2ml centrifuge tube, add 700μl lysis buffer to the centrifuge tube, add 5μl β-mercaptoethanol, and preheat at 60°C for 10min; 2) Take 0.1-0.15g of fresh raspberry leaves, add 0.1g Vinylpyrrolidone was ground into powder under liquid nitrogen freezing conditions, quickly transferred to the centrifuge tube in step 1), vortexed for 1 min, and kept at 60°C for 30 min; 3) After the mixture in step 2) was cooled to room temperature , add 700 μl of water-saturated phenol, 12 μl of glacial acetic acid, and 140 μl of chloroform in sequence, and shake vigorously for 2 minutes; 4) Centrifuge at 16,000×g for 10 minutes at 4°C; 5) Transfer 500 μl of the supernatant from step 4) to another centrifuge tube, add 1250 μl -20°C pre-cooled absolute ethanol to the tube, mix upside down and stand at -20°C for 30 minutes, centrifuge at 16000×g for 10 minutes at 4°C, discard the supernatant, and keep the precipitate; 6 ) washing the precipitate obtained by centrifugation in step 5) ...
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