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

Inactive Publication Date: 2007-01-31
JIERUI BIOENG SHANGHAI
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  • Summary
  • Abstract
  • Description
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Problems solved by technology

Since the method of Han et al. needs to use a small amount of 5M guan

Method used

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Embodiment Construction

[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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Abstract

The present invention relates to an one-step method ribonucleic acid extraction reagent EZRNA. Its formula includes (by 1 L total amount) guanidinium isothiocyanate 3-5 M, sodium citrate 18-25 M, sodium laurate (by wt%) 0.3-0.6%, 2-mercaptoethanol 0.6-1.2 M and balanced acidic phenol (by volume%) 40-55%. Besides, said invention also provides the concrete steps of the invented one-step method ribonucleic acid extraction process by using said reagent.

Description

technical field [0001] The one-step ribonucleic acid extraction reagent of the present invention relates to a reagent for one-step extraction of ribonucleic acid (hereinafter referred to as RNA) and an extraction process thereof. Background technique [0002] In order to quickly isolate complete RNA from cells, many methods use strong denaturants to lyse cells while dissolving and denaturing endogenous RNases. This method was officially published by Han (1987) et al. and Chirgwin (1979) et al. Articles are detailed. Since the method of Han et al. needs to use a small amount of 5M guanidine thiocyanate to dissolve RNA continuously, it is relatively cumbersome. This method is almost replaced by the one-step method of Chomczynski and Sacchi for this reason, and this method does not need ultracentrifugation, thereby makes many samples can be operated and speed up, and the present invention is the RNA extraction reagent developed by oneself on this basis. Currently, the RNA ext...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 赫英俊李正友邵永胜肖爱军
Owner JIERUI BIOENG SHANGHAI
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