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Nucleic acid extraction method and kit for nucleic acid extraction

An extraction method and kit technology, applied in the direction of DNA preparation, recombinant DNA technology, etc., can solve the problems of nucleic acid protein pollution, nucleic acid extraction obstacles, increase the nucleic acid extraction process, etc., and achieve the effect of good nucleic acid quality and good repeatability

Pending Publication Date: 2021-06-18
SOUTH UNIVERSITY OF SCIENCE AND TECHNOLOGY OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In the existing nucleic acid extraction technology, proteinase K is usually used to digest protein to release nucleic acid. If proteinase K is not added, the release of nucleic acid is insufficient, and protein contamination is prone to occur in the extracted nucleic acid.
Proteinase K is a highly active serine protease, but there is no unified standard for the application conditions of proteinase K. Various solvents and conditions in the extraction reagent may affect the activity of proteinase K, resulting in obstacles to nucleic acid extraction.
Moreover, the reaction temperature of proteinase K is 50-65°C, and heat treatment is required, which increases the process of nucleic acid extraction, and the efficiency and stability of nucleic acid extraction are low

Method used

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  • Nucleic acid extraction method and kit for nucleic acid extraction
  • Nucleic acid extraction method and kit for nucleic acid extraction
  • Nucleic acid extraction method and kit for nucleic acid extraction

Examples

Experimental program
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Embodiment 1

[0049] The embodiment of the present invention provides a kit for nucleic acid extraction, the components of which are shown in Table 1.

[0050] Table 1 Kits

[0051]

[0052] Remarks: The nano-magnetic bead solution is a brown solution. Before using the nano-magnetic bead solution, the nano-magnetic beads should be resuspended and mixed by vortexing. Other solutions should be colorless and clear liquids at room temperature (15-30°C). If a precipitate is observed, the reagent should be heated at 37°C until the precipitate dissolves.

[0053] In addition, the validity period of the kit provided by the present invention is 12 months, and the amount of reagents recorded in Table 1 is for reference, and in other embodiments, it can be adjusted according to actual needs.

[0054] Wherein, the lysate includes: guanidine isothiocyanate (strongly ionic chaotropic salt), Brij25 (surfactant) and absolute ethanol (alcoholic solution).

[0055] The washing solution included absolut...

Embodiment 2

[0088] The present invention provides a nucleic acid extraction method and its kit, which are roughly the same as in Example 1, except that the lysate is different. The strongly ionic chaotropic salt in the lysate provided in this example is guanidine hydrochloride.

Embodiment 3

[0090] The present invention provides a nucleic acid extraction method and its kit, which are roughly the same as in Example 1, except that the lysate is different. The strongly ionic chaotropic salt in the lysate provided in this example is sodium iodide.

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Abstract

The invention discloses a nucleic acid extraction method and a kit for nucleic acid extraction, and relates to the technical field of nucleic acid extraction.The extraction method comprises the steps that a sample is split through a lysis solution, and the lysis solution comprises strong ionic chaotropic salt with the working concentration being 0.5-6 M, a surfactant with the working concentration being 0.5-10 v / v% and an alcoholic solution with the working concentration being 10-50 v / v%; the lysate can effectively and stably lyse a sample under the condition that protease K is not needed, heating is not needed in the lysing process, effective extraction of nucleic acid can be completed under the normal-temperature reaction condition, in addition, magnetic beads are added in the sample lysing process in the extraction method, lysing and combining are completed in one step, the extraction quality is guaranteed, and meanwhile the extraction efficiency is improved, and the operation process is simplified.

Description

technical field [0001] The invention relates to the technical field of nucleic acid extraction, in particular to a nucleic acid extraction method and a nucleic acid extraction kit. Background technique [0002] Nucleic acid is the carrier of genetic information, a very important biological information molecule, and the main object of molecular biology research. Therefore, the quality of nucleic acid extraction will directly affect the success or failure of the experiment. [0003] Nucleic acid is always combined with various proteins in cells. The separation of nucleic acids mainly refers to the separation of nucleic acids from biological macromolecules such as proteins, polysaccharides, and fats. When isolating nucleic acid, on the one hand, it is necessary to ensure the integrity of the nucleic acid structure, because a complete primary structure is the basic requirement for the study of nucleic acid structure and function; on the other hand, it is also necessary to elimi...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1013
Inventor 刘莹张博宋昆李毅邵攀霖程立傅暘杨烽王光会
Owner SOUTH UNIVERSITY OF SCIENCE AND TECHNOLOGY OF CHINA
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