Preserved lysate and sample nucleic acid rapid extraction method based on same

Abstract

The invention relates to a preserved lysate and a sample nucleic acid rapid extraction method based on the same. The preserved lysate comprises the following components in percentage by volume: 5-30% of absolute ethyl alcohol and 70-95% of a mixed aqueous solution, wherein based on the volume of the preserved lysate, the mixed aqueous solution comprises the following solutes in concentration: 0.5-1mol / L of ammonium chloride, 10-50mmol / L of trihydroxymethyl aminomethane hydrochloride, 0.4-2mol / L of guanidine thiocyanate, 0.4-1mol / L of sodium chloride, 10-50mmol / L of ethylenediamine tetraacetic acid and 0.1-2mmol / L of tris- (2-formylethyl) phosphine hydrochloride. The method comprises the following steps of: (a) putting a sample into a closed container filled with nano beads and the preserved lysate; (b) lysing the sample; and (c) incubating at room temperature, and performing centrifugal washing. By virtue of the prepared reagents, full lysis of cells and nucleic acid protection are realized by combining chemical and physical methods.

Description

technical field [0001] The invention belongs to the technical field of nucleic acid extraction, and in particular relates to a preserved lysate and a method for rapidly extracting sample nucleic acid based on it. Background technique [0002] For molecular detection based on nucleic acid amplification and sequencing technology, how to preserve and obtain high-quality nucleic acid in different scenarios is a necessary factor to ensure the success of downstream nucleic acid analysis. Nucleic acid in biological samples will degrade rapidly at room temperature, especially in the field without low temperature storage conditions, the degradation of nucleic acid will be particularly serious. In the case of lack of effective sample size, it is particularly important to effectively obtain high-quality nucleic acid methods. In the process of processing nucleic acids, nucleases in the environment can degrade nucleic acids; on the other hand, in the process of processing infectious biol...

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Problems solved by technology

[0005] However, the existing nucleic acid extraction technology mainly has the following defects: (1) It cannot be used in conjunction with clinical needs: clinical samples usually need to use transport fluid or clinical sample storage fluid; transport fluid or storage fluid will inhibit downstream molecular detection. required reaction; and the transport solution or storage solution will greatly dilute the concentration of the sample, thereby seriously affecting the detection limit and causing false negatives; (2) the existing kits usually cannot completely extract the sampled sample, but only the sampled sample Part of it is used for nucleic acid extraction; (3) Most of the lysate reagents used are limited to the extraction function and have limited protection for nucleic acids; (4) Most kits are only applicable to specific cells: for example, only applicable to Bacterial, only applicable to plant cells, only applicable to animal cells, etc.

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