One step method ribonucleic acid extraction agent
A technology of ribonucleic acid and reagents, applied in the field of one-step extraction of ribonucleic acid reagents and its extraction process, can solve cumbersome problems, achieve low prices and reduce production costs
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[0029] The one-step method RNA extraction reagent of the present invention, specific formula (in 1L total amount) is as follows: guanidine isothiocyanate 3~5M, sodium citrate 18~25M, sodium laurosilicate is calculated as 0.3~0.6% by weight percentage , 2-mercaptoethanol 0.6-1.2M, equilibrium acidic phenol is 40-55% by volume percentage.
[0030] The extraction steps are as follows:
[0031] Grind the tissue into a powder in liquid nitrogen, and transfer the powder to a sterile 1.5ml centrifuge tube before the liquid nitrogen has completely evaporated. After the cells were counted, they were directly added to the centrifuge tube, centrifuged at 5,000 rpm (rev / min) at room temperature, and the supernatant was removed.
[0032] Add RNA extraction reagent, per 100mg tissue or 5×10 6 Add 1.0ml of RNA extraction reagent to each cell, and repeatedly pump and mix.
[0033] Add glycogen to make the final concentration 250μg / ml, shake vigorously or homogenize with a homogenizer. Asp...
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