Method for identifying sex of ancient human remains with very low DNA content

An identification method and gender technology, applied in the field of gender identification of ancient human remains, can solve the problems of low ancient DNA content, serious degradation, misreading, etc., and achieve the effects of improving detection sensitivity, reducing operation steps, and shortening experimental time.

Active Publication Date: 2019-07-26
XI AN JIAOTONG UNIV
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Problems solved by technology

However, for remains with poor preservation, the ancient DNA content is extremely low and the degradation is serious, and it is difficult to reach the detection limit of the ancient DNA band by electrophoresis. The gene on the X or Y chromosome may only amplify a band, according to the gender analysis method , women usually have one belt, while men have two belts, so there will be misreadings, which will affect the authenticity of the results and greatly reduce the reliability of gender results

Method used

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  • Method for identifying sex of ancient human remains with very low DNA content
  • Method for identifying sex of ancient human remains with very low DNA content
  • Method for identifying sex of ancient human remains with very low DNA content

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Embodiment 1

[0054] 1. Experimental samples: 5,000 years ago, the remains of ancient humans at the Wuzhuang Guocheng site in the Neolithic Age were used as the research object. The standards for pollution prevention were strictly followed when collecting samples. The collectors used disposable non-polluting masks, hats and gloves and non-polluting appliances. Three samples AH1-2, AH1-3, AH1-13.

[0055] 2. Experimental methods and results

[0056] Step 1, ancient DNA extraction from the Wuzhuang Guocheng ruins: Weigh 500mg of bone powder, add 1ml of 10% SDS; 4ml of EDTA (PH 8.0) and 100μl of 10mg / ml proteinase K, shake overnight at 37°C; DNA after overnight The extracted samples were transferred to an air bath for shaking, continued shaking and incubating at 55°C for 6h, and then placed at 95°C for 5min to inactivate excess proteinase K. After the lysate was centrifuged at 7500r / min for 8min, 800μl of the supernatant was added to an ultrafiltration centrifuge tube and centrifuged at 6800...

Embodiment 2

[0077] 1. Experimental sample: Lishan Fenglin Xiangxi Site, a building with a patio. A long slope tomb that passes through the cave. Two human bones were found in this tomb, two of which had the same head orientation and were located within the coffin marks. , and the head orientation is opposite to the above two, the specific burial style is unclear.

[0078] 2. Experimental methods and results

[0079] Step 1. Ancient DNA extraction from Lishan Fenglin Xiangxi site: Weigh 500mg of bone powder, add 1ml of 10% SDS; 4ml of EDTA (pH 8.0) and 100μl of 10mg / ml proteinase K, shake overnight at 37°C; The DNA extraction sample was transferred to an air bath for shaking, continued shaking and incubating at 55°C for 6h, and then placed at 95°C for 5min to inactivate excess proteinase K. After the lysate was centrifuged at 7500r / min for 8min, 800μl of the supernatant was added to an ultrafiltration centrifuge tube, centrifuged at 6800r / min, and the lysate was concentrated to 100μl. A...

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Abstract

A method for identifying the sex of ancient human remains with a very low DNA content includes first identifying extracted DNA; then setting two pairs of sex identification sites, and marking the sexidentification sites as sex sites A and B, wherein the sex site A is an enamel protein gene, the sex site B is a conserved region on X and Y chromosomes; and amplifying and identifying the sex sites Aand B respectively by means of a real-time fluorescent quantitative PCR-MGB probe method. Two sex amplification genes are combined, two sex gene identification results can complement and verify eachother, and the reliability of the ancient human remains sex identification results is greatly increased. The amount of a ancient DNA template used in the method is less, and the whole process of two sex gene identification can be quantified by the template as long as 1.5 [mu]l is adopted; compared with application of conventional PCR amplification and gel electrophoresis verification steps, application of a real-time fluorescent quantitative PCR technology shortens experiment time and reduces the operation steps, and the detection sensitivity and efficiency are improved.

Description

technical field [0001] The invention relates to the field of molecular archaeology, in particular to a method for gender identification of ancient human remains with extremely low DNA content. Background technique [0002] Modern Chinese archaeology uses a group of utensil types with clear characteristics and often accompanying them as symbols to distinguish different cultures. This kind of archaeological concept just excludes the people who created history and culture. In fact, whether it is ancient or modern, human beings will always be The main body of social activities, human society should be most concerned about human beings themselves. Therefore, the rise of ancient DNA molecular anthropology in our country is precisely to break this single, morphologically corresponding cultural qualitative research model, and to inject all the information of ancient human life and material remains into the process of human history research, thereby pulling human history research to ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6879C12Q1/6851
CPCC12Q1/6851C12Q1/6879C12Q1/6888C12Q2531/113C12Q2563/107C12Q2545/114
Inventor 张虎勤赵静吴晓明刘晓刚杜建强林松胡曦
Owner XI AN JIAOTONG UNIV
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