Preserved lysate and sample nucleic acid rapid extraction method based on same
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An extraction method and lysate technology, applied in the field of preservation of lysate and rapid extraction of nucleic acid from samples based thereon, can solve problems such as inability to cooperate with clinical needs, the kit cannot completely extract samples, and the limit of detection is affected.
Inactive Publication Date: 2021-08-06
常州艾立贝医疗科技有限公司 +1
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[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 detecti
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Embodiment 1
[0032] The present embodiment provides a kind of preserved lysate, and it comprises the component of following volume content:
[0033] Absolute ethanol 5%;
[0034] Mixed aqueous solution 95%;
[0035] Based on the volume of the preserved lysate, the mixed aqueous solution contains the following concentrations of solutes:
[0036]
[0037]
[0038] The sample nucleic acid quick extraction method that adopts above-mentioned preserved lysate to carry out, it comprises the following steps:
[0039] (a) Place the sample in an airtight container containing the nanobeads and the preserved lysate; specifically, collect the sample with a cotton swab, and then insert the cotton swab into the preservation tube containing the nanobeads and the preserved lysate In the middle, screw the cap tightly (the sampling cotton swab, preservation tube, preservation lysate and nano-beads can be combined to realize module integration);
[0040] (b) Cleavage the sample, then transfer it to a...
Embodiment 2
[0044]This embodiment provides a kind of preserved lysate, which is basically consistent with that in Example 1, except that:
[0045] Based on the volume of the preserved lysate, the mixed aqueous solution contains the following concentrations of solutes:
[0046]
Embodiment 3
[0048] This embodiment provides a kind of preserved lysate, which is basically consistent with that in Example 1, except that:
[0049] Based on the volume of the preserved lysate, the mixed aqueous solution contains the following concentrations of solutes:
[0050]
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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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