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A method for sexing ancient human remains with extremely 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 effect of improving detection sensitivity, reducing operation steps, and shortening experiment time

Active Publication Date: 2021-02-02
XI AN JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • A method for sexing ancient human remains with extremely low DNA content
  • A method for sexing ancient human remains with extremely low DNA content
  • A method for sexing ancient human remains with extremely low DNA content

Examples

Experimental program
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Effect test

Embodiment 1

[0054]1. Experimental samples: The remains of ancient humans at the Wuzhuang Guoyan site in the Neolithic age were taken as the research object 5000 years ago. The samples were collected in strict compliance with pollution prevention standards, and the collectors used disposable non-polluting masks, hats and gloves And pollution-free appliances. Three samples AH1-2, AH1-3, AH1-13.

[0055]2. Experimental methods and results

[0056]Step 1. Extraction of ancient DNA from Wuzhuang Guoyan site: Weigh 500mg bone meal, add 1ml 10% SDS; 4ml EDTA (PH 8.0) and 100μl 10mg / ml proteinase K, shake overnight at 37℃; DNA after overnight The extracted samples were transferred to the air bath and shaken, and the shaking was continued at 55°C for 6 hours, and then placed at 95°C for 5 minutes to inactivate excess proteinase K. After centrifuging the lysate at 7500r / min for 8min, add 800μl of the supernatant to the ultrafiltration centrifuge tube and centrifuge at 6800r / min to concentrate the lysate to 100...

Embodiment 2

[0077]1. Experimental sample: The Xiangxi Ruins of Fenglin, Lishan, a well with a patio. There are two human bones found in the tomb with a long slope through the tomb. Two of them have the same head and are located within the coffin scar. One of them is buried with straight limbs and the other is buried with curved limbs. , And the head is opposite to the above two, the specific burial style is unclear.

[0078]2. Experimental methods and results

[0079]Step 1. Extraction of ancient DNA from the Xiangxi site of Fenglin, Lishan: Weigh 500mg bone meal, add 1ml of 10% SDS; 4ml of EDTA (PH 8.0) and 100μl of 10mg / ml proteinase K, shake overnight at 37℃; after overnight The extracted DNA samples were transferred to the air bath for shaking, and the shaking was continued at 55°C for 6 hours, and then placed at 95°C for 5 minutes to inactivate excess proteinase K. After centrifuging the lysate at 7500r / min for 8min, take 800μl of the supernatant and add it to the ultrafiltration centrifuge tube...

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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 sex identification of ancient human remains with extremely low DNA content.Background technique[0002]Modern Chinese archaeology uses a group of artifacts with clear characteristics that often appear as a symbol to distinguish different cultures. This archaeological concept excludes those who create history and culture. In fact, whether ancient or modern, human beings will always be The subject of social activities, what human society cares most about should be human beings. Therefore, the rise of ancient DNA molecular anthropology in my country is precisely to break this single, morphologically corresponding cultural qualitative research model, and to inject all the information of ancient human life material remains into the process of human history research, thereby pulling human history research On the human-centered research track, anthropology, archaeology and life sci...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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