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Primer composition, kit and method for composite identification of polymorphic genetic marker of industrial cannabis sativa

A primer composition and polymorphic inheritance technology, which is applied in the field of primer composition for compound identification of polymorphic genetic markers in industrial hemp, and achieves the effects of good specificity, strong stability and high sensitivity

Inactive Publication Date: 2021-06-25
ACADEMY OF FORENSIC SCIENCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Foreign research on the STR compound amplification system of industrial hemp is in its infancy, and there is almost no research on it in China

Method used

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  • Primer composition, kit and method for composite identification of polymorphic genetic marker of industrial cannabis sativa
  • Primer composition, kit and method for composite identification of polymorphic genetic marker of industrial cannabis sativa
  • Primer composition, kit and method for composite identification of polymorphic genetic marker of industrial cannabis sativa

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Experimental program
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Embodiment 1

[0048] This embodiment provides a method for compound identification of polymorphic genetic markers in industrial hemp, comprising the following steps:

[0049] 1. Screening of industrial hemp STR loci suitable for forensic applications;

[0050] Based on literature reports and gene databases, search for the STR loci that have been developed so far, but not all of these STR loci are suitable for the research on the multiple amplification system of industrial hemp. Screen the polymorphic STR loci and eliminate the loci with dinucleotide repeats. Finally, 17 optimal autosomal STR loci and 2 sex identification loci were selected for kit development, and the loci were D02-CANN1, C11-CANN1, DM029, DM016, 4910, B01-CANN1, E07- CANN1, 9269, B05-CANN1, H06-CANN2, 5159, nH09, ANUCS 501, CS1, ANUCS 305, 3735, ANUCS 302, 1528 and 9043. Among them, the gender identification site DM029 is related to the X chromosome, and all samples have a single peak, and the sex identification site DM0...

Embodiment 2

[0069] In this example, the method provided in Example 1 is used to detect the DNA of the industrial hemp sample.

[0070]Different tissues (flowers, stems, leaves, and seeds) were collected from a sample of the genus Hemp for DNA extraction and quantification. The kit constructed in Example 1 was used to perform multiplex amplification of 19 genetic markers on the above DNA samples. All samples obtained effective amplification products on genetic markers.

Embodiment 3

[0072] According to the requirements of SWGDAM, this embodiment performs forensic verification (identity, sensitivity, species specificity and forensic parameters calculation) on the method provided in Example 1, and then judges the superiority of the method. The specific operations are as follows:

[0073] (1) Identity study: Quantify the genomic DNA of flowers, leaves and stems of the same plant to 1 ng / μL, and use the method provided in Example 1 to detect them respectively.

[0074] The results show (such as Figure 4 shown), the method constructed in Example 1 has good tissue identity: flowers, leaves and stems of the same plant have the same genotype.

[0075] (2) Sensitivity study: the industrial hemp genomic DNA was diluted to 2ng / μL, 1ng / μL, 0.5ng / μL, 0.25ng / μL, 0.125ng / μL, 0.0625ng / μL, 0.03125ng / μL and 0.015625 ng / μL, using the method provided in Example 1 to detect the industrial hemp genomic DNA at the above concentrations, and repeat the detection 3 times for ea...

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Abstract

The invention provides a primer composition, a kit and a method capable of detecting 17 autosomal STR gene loci and two sex identification loci of an industrial cannabis sample in parallel. The 17 autosomal STR gene loci comprise D02-CANN1, C11-CANN1, 4910, B01-CANN1, E07-CANN1, 9269, B05-CANN1, H06-CANN2, 5159, nH09, ANUCS 501, CS1, ANUCS 305, 3735, ANUCS 302, 152 and 9043, and the two sex identification loci are DM029 and DM016; and the primer composition comprises the sequences as shown in SEQ ID NO. 1 to 38. The primer composition, the kit and the method provided by the invention have the characteristics of high sensitivity, good specificity, strong stability, high accuracy and the like, and the primer composition, the kit and the method can be used for detecting 17 autosomal STR gene loci and 2 sex identification loci of industrial cannabis samples in parallel; a brand new detection means is provided for sex identification, individual identification, source inference and STR database establishment in forensic medicine research of industrial cannabis sativa in China.

Description

technical field [0001] The invention relates to the technical field of molecular identification, in particular to a primer composition, a kit and a method for compound identification of polymorphic genetic markers in industrial hemp. Background technique [0002] Industrial hemp is an annual dioecious herbaceous plant belonging to the genus Cannabis of the family Cannabinaceae. It is mainly distributed in Nepal, India, China and other regions in Europe, Africa and Asia. It is the oldest cultivated crop on earth. one. Industrial hemp fiber is a good raw material for textile and papermaking. The industrial hemp contained in the plant is widely used in the field of medicine and health. The seeds are rich in protein, amino acids, unsaturated fatty acids and trace elements that are easily absorbed by the human body. It has become an important economic crop in many countries. [0003] Cytological studies have shown that industrial hemp is a diploid plant containing 20 chromosome...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6879C12Q1/686C12N15/11
CPCC12Q1/6895C12Q1/6879C12Q1/686C12Q2600/156C12Q2525/151
Inventor 张素华李成涛夏若成于欢陶瑞旸陈安琪刘希玲阙庭志林源赵珍敏
Owner ACADEMY OF FORENSIC SCIENCE
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