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Free DNA extraction kit and application thereof

A kit and inhibitor technology, applied in the field of cell-free DNA extraction, can solve the problems of high equipment price, large volume, and high cost

Inactive Publication Date: 2016-08-17
SHANGHAI KEHUA BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These platforms also have the disadvantages of large size, complex structure, high equipment price, and high cost per operation, which cannot meet the needs of existing users for automatic processing of cell-free DNA.

Method used

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  • Free DNA extraction kit and application thereof
  • Free DNA extraction kit and application thereof
  • Free DNA extraction kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Using a fully automatic cell-free DNA extraction system and supporting cell-free DNA extraction kits, the simulated plasma cell-free DNA samples were extracted, and Tiangen plasma cell-free DNA extraction kits (catalog number DP339, Tiangen Biochemical Technology (Beijing) Co., Ltd., Beijing ) were compared to compare the extraction efficiencies of the two methods on simulated plasma cell-free DNA samples.

[0070] Wherein, the free DNA extraction kit of the present invention consists of:

[0071] Lysis solution (50% guanidine isothiocyanate, 10% Triton-100, 1% sodium citrate);

[0072] Washing solution A (30% guanidine isothiocyanate, 5% Triton-100, 1% sodium citrate, 30% ethanol);

[0073] Washing solution B (0.1mol / L, Tris-HCL buffer solution with a pH value of 4.5);

[0074] Eluent (0.05mol / L, Tris-HCL buffer solution with a pH value of 8.5);

[0075] Magnetic bead solution (silane magnetic beads, the content is 30%);

[0076] De-inhibitor (30mg / ml proteinase K)...

Embodiment 2

[0083]Use the automatic free DNA extraction system and supporting free DNA extraction kit for anti-pollution test.

[0084] Simulated plasma cell-free DNA sample preparation: the artificially synthesized 161bp DNA fragment was diluted with human plasma to a certain number of times, so that the simulated DNA concentration of the diluted sample reached 10 10 copies / ml or so.

[0085] Anti-pollution test plan: the simulated plasma free DNA sample is used as a positive sample, and the negative plasma is used as a negative sample, arranged at intervals, a total of 96 samples are tested, and 48 negative samples are required not to be contaminated, and the fluorescent quantitative PCR method is used for negative and positive judgments .

[0086] The test results showed that the automatic free DNA extraction system tested 96 samples separated by yin and yang, and the test results of 48 negative samples were all negative, and no contamination occurred during the extraction process. I...

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Abstract

The invention discloses a free DNA extraction kit. The kit comprises a lysis solution, a scrubbing solution A, a scrubbing solution B, an inhibitor, magnetic beads and eluent, wherein the concentration of alcohol in the scrubbing solution B can be lower than 1%, or the scrubbing solution B is completely free of alcohol and surfactant. The invention further provides a full-automatic nucleic acid extraction system which is used in cooperation with the kit or comprises or integrates the kit. The kit is particularly suitable for extracting short-fragment DNA like plasma free DNA, does not adversely affect following PCR reaction and has the advantages of being safe and easy to use, fast, low in cost and the like.

Description

technical field [0001] The invention relates to the field of extraction of genetic material. Specifically, the present invention relates to kits for free DNA extraction and applications thereof. Background technique [0002] Cell-free DNA, also known as circulating nucleic acid, refers to the endogenous DNA that is free in the extracellular space in circulating blood, and can exist in the form of single-stranded or double-stranded DNA, most of which are double-stranded DNA in the form of nucleoproteins strand DNA. Cell-free DNA fragments are small, generally ranging in length from tens to hundreds of bp. Studies in recent years have found that cell-free DNA in human circulating blood contains very important genetic information. The use of cell-free DNA high-throughput sequencing technology can effectively carry out prenatal screening and detection of diseases such as tumors, such as Down syndrome pregnancy. For mid-term screening, the traditional method generally needs to...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1013C12Q2563/143C12Q2563/149C12Q2527/125
Inventor 陈威蒋淼
Owner SHANGHAI KEHUA BIO ENG
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