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Method for extracting ancient human remains matter DNA

An extraction method and material technology, which is applied in the field of DNA extraction from ancient human remains, can solve the problems of false positives, inability to effectively remove lysate proteins, organic reagents, and low success rate, and achieve the effect of increasing the content

Inactive Publication Date: 2006-08-23
XI AN JIAOTONG UNIV
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Problems solved by technology

[0026] However, the success rate of the DNA fragment extracts obtained by the above-mentioned extraction schemes is lower than 30% (Herrmann.Betal.1994; MacHughDeetal.2000). The reason for this is that the main defect is that the residual protein in the lysate cannot be effectively removed. , organic reagents, etc., cannot fully recover the DNA fragments in the lysate; and these extraction schemes perform generally in controlling endogenous and exogenous contamination, and are prone to false positives during the extraction process

Method used

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  • Method for extracting ancient human remains matter DNA

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Effect test

Embodiment 1

[0045] According to the method of the present invention, the bone sample is treated with plasma, etc., so that the organic pollutants on the inner surface of the bone sample are decomposed, the surface and inner surface pollution of the bone sample are quickly removed, and then the DNA is extracted; specifically, the following steps are included:

[0046] 1. For samples of ancient humans, select bones and teeth, polish the surface to 5mm, and then treat the surface with hypochlorous acid.

[0047] 2. Perform 30min ultraviolet (λ=254nm) irradiation and wash twice with 70% ethanol to remove exogenous microbial DNA contamination.

[0048] 3. Place the sample in the plasma treatment reactor, and the vacuum degree of the system reaches 10 -1 Pa, the inside of the plasma processing reactor includes a sample stage, an external electrode (AI), and a vacuum gauge, and the plasma processing reactor is connected with a radio frequency power generator, a vacuum pump, an Ar plasma ( figur...

Embodiment 2

[0061] The difference between this embodiment and Embodiment 1 is that the discharge time of the radio frequency power generator is 10 minutes, and the rest are the same as Embodiment 1, and the purpose of Embodiment 1 can be achieved as well.

Embodiment 3

[0063] The difference between this embodiment and Embodiment 1 is that the discharge time of the radio frequency power generator is 1 min, and the rest are the same as Embodiment 1, and the purpose of Embodiment 1 can also be achieved.

[0064] Many examples can be cited in the above-mentioned embodiments, and it has been proved by many experiments that the discharge time of the radio frequency power generator is 1min-10min, which can better achieve the purpose of the present invention.

[0065] scheme and comparison

[0066] It can be seen from the above table that the method of the present invention has obvious advantages in the concentration (λ=260) and purity (A260 / A280) of the extract, and the success rate of extraction far exceeds that of the traditional extraction method.

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Abstract

The present invention relates to method of extracting ancient human remains matter DNA. The method includes first treating skeleton sample with plasma to decompose the surface pollutant and subsequent DNA extraction. The DNA extraction includes the steps of polishing skeleton sample, hypochloric acid treatment, ultraviolet irradiation, alcohol solution washing to eliminate foreign microbe contamination, Ar / O2 (9:1) plasma treatment, grinding in liquid nitrogen to obtain powder sample; digesting and cracking sample, phenol-chloroform extraction to eliminate digested organic molecule, adsorption with silica adsorption column, and filtering in 3 KDa ultrafiltering centrifugation tube to obtain ancient human remains matter DNA. The method of the present invention can eliminate surface pollutant fast effectively, and has high ancient human remains matter DNA extracting success rate and high template content.

Description

technical field [0001] The invention relates to the extraction and purification of ancient human DNA, in particular to a method for extracting ancient human DNA, which can prevent endogenous and exogenous pollution and improve the success rate of extraction. Background technique [0002] At present, ancient DNA extraction methods are mainly based on some extraction methods in forensic science. These extraction schemes generally first remove the exogenous DNA on the sample surface by polishing the sample surface or sawing off the sample surface by 1-2mm with a wire saw, ultraviolet irradiation, and washing with 70% ethanol. contamination, and then grind the sample into powder in liquid nitrogen for pre-sample treatment. Specifically, there are mainly three different extraction schemes: proteinase K / phenol-chloroform method, silica method and guanidine isothiocyanate-silica method. [0003] 1. The step of proteinase K / phenol-chloroform method (ThomasWK...

Claims

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Application Information

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IPC IPC(8): C07K1/14
Inventor 张虎勤张金刘芳娥葛宇
Owner XI AN JIAOTONG UNIV
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