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Kit and method for extracting circulating DNA in plasma

An extraction method and plasma technology, applied in the field of DNA extraction, can solve the problems of high price, long extraction time, cumbersome methods, etc., and achieve the effects of preventing dust pollution, simplifying extraction steps and operations, and prolonging extraction time.

Active Publication Date: 2019-02-19
北京爱普拜生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In view of the shortcomings of the above plasma circulating DNA extraction methods, such as long extraction time, cumbersome methods, low extraction purity, low extraction yield and high price, one of the purposes of the present invention is to provide a kit for extracting circulating DNA from plasma, Using this kit to extract circulating DNA in plasma, the operation is fast and simple, the cost is low, and the content and purity are high

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  • Kit and method for extracting circulating DNA in plasma
  • Kit and method for extracting circulating DNA in plasma
  • Kit and method for extracting circulating DNA in plasma

Examples

Experimental program
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Effect test

Embodiment 1

[0059] Embodiment 1: kit of the present invention

[0060] The kit for extracting circulating DNA from plasma samples comprises:

[0061] DNA one-step reaction binding buffer DRB Buffer, cleaning solution DRBW Buffer, eluent DRBE Buffer, independent aseptic packaging cycle DNA purification extension tube SSPET. The DNA one-step reaction binding buffer DRB Buffer includes 3-5 mol / L urea, 0.5-3% sodium lauryl sarcosinate by mass volume fraction, 10-60% Tris saturated phenol by volume fraction, and The fraction is 0.5-5% Tween20, the volume fraction is 5-32% isopropanol, and the pH value is 5.9-6.5; the cleaning liquid DRBW Buffer contains 0.1-5 mmol / L, a Tris buffer solution with a pH of 7.0, 5 ~50mmol / L sodium chloride, the volume fraction is 55~85% absolute ethanol; the eluent DRBEBuffer contains 0.5~5mmol / L, Tris-HCl buffer solution with a pH of 8.5; the independent sterile packaging The volume specification of the circulating DNA purification extension tube is 10-50 ml;

...

Embodiment 2

[0068] Using the kit described in Example 1, the circulating DNA in the plasma sample was extracted, and parallel comparison was performed with the extraction method of the commonly used kit.

[0069] 1. Extraction reagent

[0070] DNA one-step reaction binding buffer DRB Buffer: 4.5mol / L urea, the concentration is 1.5% sodium lauryl sarcosinate by mass volume fraction, 40% Tris saturated phenol by volume fraction, and 2% by volume fraction Tween 20, volume fraction is 20% isopropanol, pH value is 6.2;

[0071] Cleaning solution DRBW Buffer: 2mmol / L, Tris buffer solution with pH 7.0, sodium chloride with a concentration of 10mmol / L, and absolute ethanol with a volume fraction of 80%;

[0072] Eluent DRBE Buffer: 2mmol / L, Tris-HCl buffer with pH 8.5;

[0073] Individually sterile packaged circular DNA purification extension tube SSPET.

[0074] 2. Extraction method

[0075] (1) Take 1 ml of fresh plasma and centrifuge at 14,000 rpm for 10 minutes at 5°C to remove impurities...

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Abstract

The invention provides a kit and a method for extracting circulating DNA from plasma. The kit provided by the invention comprises a DNA one-step reaction binding buffer solution, a washing solution, an eluent, and an independent aseptic packaged circulating DNA purification extension tube used in combination with a DNA binding column. The kit and the method provided by the invention can ensure that plasma samples having different starting volumes pass through the DNA binding column at a time, can effectively avoid cross-contamination between samples while rapidly and efficiently extracting high-quality circulating DNA, and have obvious advantages in comparison with common kits and methods.

Description

technical field [0001] The invention relates to DNA extraction and belongs to the field of nucleic acid purification, in particular to a kit and method for simply and efficiently extracting circulating DNA from blood plasma. Background technique [0002] Blood circulating DNA is also called blood cell-free DNA. It refers to a small length of endogenous DNA that is released into the blood by human cells, frees outside the cells and circulates in the blood. The circulating DNA secreted by cancer tumor cells into the blood is also called circulating tumor DNA ( cell-tumor nucleic acid, ctDNA), is generally believed to be derived from the active secretion of tumor cells and the passive release of apoptotic cells. The value of circulating DNA in the peripheral blood of cancer patients in the diagnosis, differential diagnosis, and prognosis evaluation of various tumors has gradually been recognized. Up to now, it has been confirmed in the research of various cancers including me...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 于祥春冯晓燕林挺王文利龚建
Owner 北京爱普拜生物技术有限公司
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