Primers, kit and method for detecting short tandem repetitive sequence

A short tandem repeat and kit technology, applied in the field of genetic engineering, can solve problems such as genetic polymorphism differences of short tandem repeat sequence loci, and achieve the effects of low cost, high accuracy and simple operation.

Active Publication Date: 2017-05-24
SHANGHAI TISSUEBANK MEDICAL LAB CO LTD +3
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  • Abstract
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AI Technical Summary

Problems solved by technology

A large number of studies have reported that the genetic polymorphisms of short tandem repeat loci are somewhat different among ethnic groups

Method used

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  • Primers, kit and method for detecting short tandem repetitive sequence
  • Primers, kit and method for detecting short tandem repetitive sequence
  • Primers, kit and method for detecting short tandem repetitive sequence

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1: Detection of 18 STR loci in human genomic DNA samples 1

[0051] In this example, magnetic beads were used to extract DNA to obtain human genomic DNA samples; PCR amplification was performed on the obtained DNA samples, and the fluorescent labels of the primers used were 6-FAM, HEX, TAMRA and ROX respectively; the obtained PCR amplification products Capillary electrophoresis detection; typing analysis. At the same time, combined with Life's STR detection kit and Sanger sequencing method to determine the types of 18 STR loci in the obtained human genomic DNA samples. details as follows.

[0052] 1. Configuration of primer composition storage solution

[0053] Prepare four sets of 5× primer storage solutions according to the formula shown in Table 3.

[0054] Table 3, the formula list of four groups of 5× primer storage solutions

[0055]

[0056]

[0057]

[0058] 2. PCR amplification

[0059] During PCR amplification, a DNA sample is amplified i...

Embodiment 2

[0070] Example 2: Detection of 18 STR loci in human genomic DNA samples 2

[0071] In this example, the magnetic bead method was used to extract DNA to obtain human genomic DNA samples; PCR amplification was performed on the obtained DNA samples, and the fluorescent labels of the primers used were 6-FAM, HEX, TAMRA and ROX respectively; the obtained PCR amplification products Capillary electrophoresis detection; typing analysis. At the same time, combined with Life's STR detection kit and Sanger sequencing method to determine the types of 18 STR loci in the obtained human genomic DNA samples. details as follows.

[0072] 1. Configuration of primer composition storage solution

[0073] According to the formula shown in Table 5, prepare 5× primer storage solution.

[0074] Formulation table of table 5, 5 × primer storage solution

[0075]

[0076]

[0077] 2. PCR amplification

[0078] During PCR amplification, 18 STR loci of a DNA sample are amplified in an amplifica...

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Abstract

The invention belongs to the gene engineering technical field, and discloses a primer combination for detecting a short tandem repetitive sequence, a kit containing the primer combination and a method for detecting the short tandem repetitive sequence. Gene loci having high China people genetic polymorphism are selected and are more suitable for China people; the number of the selected tandem repetitive units is four, five or six, the slip peak ratio can be effectively reduced, and the accuracy of test results is higher; a fluorescent labeling composite amplification system is used for amplification, and amplified products can be subjected to electrophoresis in a same capillary, to achieve detection analysis of all the gene loci at the same time; a fluorescent label with mature technology is used for fluorescent labeling, and thus the cost is low; the detection method has good specificity and can be used for qualitative analysis such as individual identification, paternity identification, population genetic studies and the like, and can also be used for qualitative analysis of detection of the chimerism rate after hematopoietic stem cell transplantation.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to primers, kits and methods for detecting short tandem repeat sequences. Background technique [0002] Short tandem repeats (short tandem repeats, STR) are a class of DNA tandem repeats widely present in the human genome, with a core sequence of 2 to 6 bases. Short tandem repeat sequences have a very high recognition rate among different individuals. According to statistics, the probability of identical 12 STR sites between unrelated individuals is 10 -14 , the probability that siblings are exactly the same is 10 -4 , is a powerful indicator to identify the DNA of different individuals, and this difference follows the Mendelian law of inheritance in the genetic process. Therefore, STR amplification detection technology is widely used in individual identification, kinship identification, population genetics research and chimerism detection and so on. [0003] With the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 郑仲征冀丽军何贵伦
Owner SHANGHAI TISSUEBANK MEDICAL LAB CO LTD
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