Kit for extracting DNA for sampling in minute quantities without wound, and method for extracting DNA

A kit and micro-quantity technology, applied in the field of biological DNA detection, can solve the problems of difficult and sensitive human sampling, and achieve the effects of reducing costs, saving resources, and improving quality and quantity

Inactive Publication Date: 2007-10-10
FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The purpose of the present invention is to propose a DNA extraction kit and a DNA extraction method that can be used for non-invasive micro or trace sampling, and can be adapted to large-scale production, so as to overcome the deficiencies of the existing DNA extraction methods and change the human body. Damage the situation of sampling, improve the quality and quantity of extraction products, and solve the difficult and sensitive problem of human sampling to a certain extent

Method used

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  • Kit for extracting DNA for sampling in minute quantities without wound, and method for extracting DNA
  • Kit for extracting DNA for sampling in minute quantities without wound, and method for extracting DNA
  • Kit for extracting DNA for sampling in minute quantities without wound, and method for extracting DNA

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1, the configuration of each component of kit

[0046] Solution composition:

[0047] (1) Lysis solution: GuSCN (guanidine isothiocyanate), EDTA, Tris-HCl, Triton X-100;

[0048] (2) Washing solution A: GUSCN (guanidine isothiocyanate), Tris-HCl;

[0049] (3) Washing solution B: 70% ethanol;

[0050] (4) washing liquid C: acetone;

[0051] (5) Eluent: Tris-Hcl, EDTA (sterilized).

Embodiment 2

[0052] Embodiment 2, human hair and saliva DNA extraction

[0053] First perform non-invasive microsampling:

[0054] (1) Hair: Pull out the hair of the experimental subject, take 1, 3, and 5 hairs respectively, cut about 0.5-1 cm from the base of the hair root with clean and dry scissors, and place them in a 1.5 mL eppendorf tube;

[0055] (2) Saliva: Use a 1.5 mL eppendorf tube to directly collect about 0.3 mL of the subject's saliva.

[0056] The specific extraction steps are as follows:

[0057] (1) Add 1mL of lysate respectively, and bathe in water at 56°C for 1h;

[0058] (2). Add 20 μl of silica gel adsorbent, mix well, and fully transpose;

[0059] (3) After sufficient centrifugation, discard the supernatant;

[0060] (4) Resuspend in 1mL washing solution A, centrifuge thoroughly, and discard the supernatant. Repeat this step once;

[0061] (5) Resuspend twice in 1mL washing solution B and centrifuge fully;

[0062](6) Resuspend once in 1mL washing solution C an...

Embodiment 3

[0066] Embodiment 3, the inspection of DNA extraction product

[0067] The extraction product obtained in Example 2 is measured at 260nm by a UV spectrophotometer, and it can be calculated that the DNA extraction amount of 1, 3, and 5 hairs is about 100ng, 300ng, and 500ng respectively, and the DNA extraction amount of 0.3mL saliva is about 100ng. It is 2.0ug, converted into the number of experiments are 10, 30, 50, 200 times. The gel electrophoresis profile of the extract is shown in Figure 4.

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Abstract

This invention discloses test kit and method for extracting DNA by noninvasive micro sampling. The test kit contains lysate solution, silica gel adsorber, cleaning solution, and eluant. The content and quality of DNA product extracted with the test kit can satisfy the following operation such as PCR and sequencing. The test kit is operated in single-tube form to prevent the loss of DNA component, and contamination during transfer. Besides, the test kit can be used together with 96 hole plate for high-flux extraction due to its in situ extraction mode. The test kit has such advantages as easy operation, low time consumption, high efficiency, and high quality and content of extracted product.

Description

technical field [0001] The invention belongs to the technical field of biological DNA detection, and in particular relates to a kit and a DNA extraction method capable of quickly, cheaply, pollution-free and non-invasively extracting human DNA. Background technique [0002] At present, most of the samples taken from the human body are blood and tissues, and the methods used are generally phenol-chloroform extraction and existing kits on the market. This sampling method is not only harmful to the human body, but also involves sensitive issues such as ethnicity and religion, and the extraction effect is not ideal. [0003] Although the traditional phenol-chloroform extraction method can extract blood DNA more efficiently, due to the use of a large number of organic reagents, not only will the extraction product contain chemical inhibitors, but it will affect the subsequent series of operations (PCR, enzyme Reaction and sequencing, etc.), and the repeated extraction and centri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/26C12Q1/68C07H21/04
Inventor 朱晔一刘启昆闫鹏荣邵敏华
Owner FUDAN UNIV
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