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Alcohol-free cleaning solution suitable for nucleic acid extraction of multiple samples and nucleic acid extraction kit

A nucleic acid extraction reagent and cleaning solution technology, which is applied in the field of alcohol-free cleaning solution and nucleic acid extraction kits, and achieves the effect of reducing impact, improving transportation safety and good stability

Active Publication Date: 2021-10-01
3D BIOMEDICINE SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It mainly solves many problems existing in the nucleic acid extraction process using ethanol cleaning in the prior art

Method used

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  • Alcohol-free cleaning solution suitable for nucleic acid extraction of multiple samples and nucleic acid extraction kit
  • Alcohol-free cleaning solution suitable for nucleic acid extraction of multiple samples and nucleic acid extraction kit
  • Alcohol-free cleaning solution suitable for nucleic acid extraction of multiple samples and nucleic acid extraction kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Extraction of oropharyngeal swab sample nucleic acid

[0043] The main steps of automatic extraction of oropharyngeal swab samples are as follows:

[0044] 1) Prepare a batch of reagents respectively according to the formula in Table 1 for future use.

[0045] Table 1

[0046]

[0047]

[0048] 2) Take a clean 96-well deep-well plate, add 700 μL of formula 1 lysate to the first column of the deep-well plate, add 900 μL of formula 1 cleaning solution A to the second column, and add 900 μL of formula 1 cleaning solution to the third column Solution B and 10 μL of 50 mg / mL silica-based magnetic beads, add 100 μL of the eluent of formula 1 to the sixth column;

[0049] 3) Add 700 μL of the lysate of formula 2 to the seventh column of the above-mentioned deep well plate, add 900 μL of cleaning solution A of formula 2 to the eighth column, add 900 μL of cleaning solution B of formula 2 and 10 μL of 100 mg / mL For silica-based magnetic beads, add 100 μL of th...

Embodiment 2

[0073] Embodiment 2 Nucleic acid extraction of the present invention is compared with similar products

[0074] The nucleic acid extraction reagent of the present invention is compared with similar products (an alcohol-free virus nucleic acid extraction kit based on the magnetic bead method, application number 201910054549.1) as follows:

[0075] 1) Formula comparison: see Table 8 and Table 9, wherein Table 8 is the formula of the extraction reagent of the present invention, and Table 9 is the formula of the extraction reagent of the comparative patent 201910054549.1.

[0076] Table 8

[0077]

[0078]

[0079] Table 9

[0080]

[0081] 2) Extraction performance comparison:

[0082] a) Reagent preparation: prepare sufficient amounts of nucleic acid extraction reagents of the present invention and comparative patents according to the above formulas;

[0083] b) Sample preparation: collect a sufficient amount of negative throat swab samples as a matrix, dilute the po...

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Abstract

The invention discloses an alcohol-free cleaning solution suitable for nucleic acid extraction of various samples and a nucleic acid extraction kit. The alcohol-free cleaning solution comprises a cleaning solution A and a cleaning solution B, the cleaning solution A comprises 30 mM to 50 mM of Tris-HCl, 15% to 30% of PEG and 2 to 3 M of sodium salt, and the cleaning solution B comprises 30 mM to 50 mM of Tris-HCl, 50 mg / mL to 100 mg / mL of silicon-based magnetic beads, 10% to 15% of PEG and 1.5 M to 2.5 M of sodium salt. The nucleic acid extraction kit provided by the invention comprises the alcohol-free cleaning solution, a lysis solution, protease K and an eluent. The nucleic acid extraction kit can be used for extracting various types of samples (oropharynx swab samples, nasopharynx swab samples, plasma, serum, sputum and saliva). By adopting the nucleic acid extraction kit, the influence of ethanol residue on downstream experiments can be reduced, the nucleic acid purity is improved, and the detection accuracy is further improved.

Description

technical field [0001] The invention belongs to the technical field of nucleic acid extraction, and in particular relates to an alcohol-free cleaning solution and a nucleic acid extraction kit suitable for nucleic acid extraction of various samples. Background technique [0002] As one of the most basic experiments in molecular experiments, nucleic acid extraction is the basis of almost all experiments. No matter the subsequent cloning, PCR, qPCR, library construction and sequencing, etc., nucleic acid is required to proceed smoothly. The extraction efficiency and purity of nucleic acid will also have a great impact on the results of downstream experiments. [0003] The main process of nucleic acid extraction is: the sample to be extracted is fully lysed in the lysate containing guanidine thiocyanate, proteinase K, dithiothreitol, trisodium citrate, Tween-20, Bridger-35 and other raw materials. The nucleic acid is released, and the released nucleic acid is combined with the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1013
Inventor 冯二环李玫杰谢正华
Owner 3D BIOMEDICINE SCI & TECH CO LTD
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