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Forensic entomology detection kit based on eight sarcophagidae mitochondrial SNP (single nucleotide polymorphisms) genetic markers

A detection kit and genetic marker technology, applied in the field of forensic medicine, can solve the problems of difficulty in achieving satisfactory accuracy, large individual hemp flies, and high cost

Inactive Publication Date: 2015-05-20
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Sixth, most species of sargassum flies are larger and more likely to attract the attention of observers
However, the current molecular identification technology of Sargassum flies still has defects
First of all, although the identification accuracy rate of full sequence analysis can reach 99.3%, it is expensive and time-consuming, and it is difficult to achieve full sequence amplification for some special samples; secondly, although the 600bp "barcode" region commonly used in the world is more effective in identification , but studies have confirmed that there is still a greater risk of using this region alone; thirdly, at present, short fragment sequences are considered to be a better choice for molecular identification, and these fragments with only about 200 bp are very easy to amplify, even if they are placed for a long time The sample can also successfully amplify the target DNA, which has obvious advantages in the identification of special samples (chrysalis, etc.), but the identification accuracy rate is only about 80%, and even multi-fragment joint analysis is still difficult to achieve a satisfactory accuracy rate

Method used

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  • Forensic entomology detection kit based on eight sarcophagidae mitochondrial SNP (single nucleotide polymorphisms) genetic markers
  • Forensic entomology detection kit based on eight sarcophagidae mitochondrial SNP (single nucleotide polymorphisms) genetic markers
  • Forensic entomology detection kit based on eight sarcophagidae mitochondrial SNP (single nucleotide polymorphisms) genetic markers

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Experimental program
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Effect test

Embodiment 1

[0060] Preparation of the kit

[0061] The Sarcophaga mitochondrial SNP composite detection kit used for detection can include the following reagents separately packaged:

[0062] a) Amplification primer pair. The amplification primer pairs shown in Table 1 were synthesized by TaKaRa Biotechnology, and the 8 pairs of amplification primers synthesized were configured to 10pM / μL with ultrapure water.

[0063] b) Pyrosequencing primers. The amplification primer pairs shown in Table 2 were synthesized by TaKaRa Biotechnology. The 8 synthesized primers were configured with ultrapure water to 10pM / μL.

[0064] c) Amplification reaction mixture. In this example, Promaga's PCR reaction mixture 2× Green Master Mix (Promerga, Madison, WI, U.S.A.).

[0065] c) Pyrosequencing purification reagents. In this example, a pyrosequencing purification kit from QIAGEN was used.

[0066] d) Pyrosequencing reagents. In this example, the PyroMark substrate Mixture pyrosequencing kit from QIAGEN was used....

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Abstract

The invention discloses a forensic entomology detection kit based on eight sarcophagidae mitochondrial SNP genetic markers. The detection kit comprises amplification primers of sequences shown in SEQ ID NO.1-SEQ ID NO.16 and sequencing primers of sequences shown in SEQ ID NO.17-SEQ ID NO.24. The kit is based on a pyrosequencing platform, so that the whole identification process only needs three hours and the identification results can be quickly obtained. The minimum length of the amplification primers of the kit is only 77bp and the maximum length does not exceed 278bp, so that the kit has advantages in detection of the common insect degrading samples on the forensic scene.

Description

Technical field [0001] The present invention belongs to the technical field of forensic medicine, and specifically relates to a forensic entomology detection kit for 8 mitochondrial SNP genetic markers used for identification of the species of Sarcophagous Sarcophagus in forensic entomology. Background technique [0002] Postmortem Interval (PMI) inference refers to the estimation and judgment of the interval between the time a corpse is discovered and the time of death. It is of great significance in providing clues to the case, delimiting the scope of investigation, analyzing the time of the crime, and determining the character of the case. PMI inference is a key link in the detection of criminal cases, and it is also a difficult and research hotspot in forensic practice. Traditionally, forensic doctors make PMI inferences based on the corpse phenomenon or the post-mortem change of the corpse's endogenous substances, but the progress of corpse corruption has seriously hindered ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6869C12Q1/6888C12Q2600/156C12Q2600/172C12Q2565/301
Inventor 郭亚东扎拉嘎白乙拉蔡继峰闫杰
Owner CENT SOUTH UNIV
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