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Detection primers, detection kits and applications based on methylation of sox10 gene in peripheral blood

A technology for detecting primers and methylation, which is applied in the field of genetic engineering, can solve the problems of less genetic detection and insufficient research, and achieve the effects of convenient operation, high medical application value, and favorable implementation

Active Publication Date: 2021-05-14
SHENGJING HOSPITAL OF CHINA MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the research on the methylation of the Sox10 gene is not sufficient, and the detection of this gene is still relatively small

Method used

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  • Detection primers, detection kits and applications based on methylation of sox10 gene in peripheral blood

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Effect test

Embodiment 1

[0037] This embodiment provides detection primers based on the methylation of Sox10 gene in peripheral blood, and the nucleotide sequences of the detection primers are shown in SEQ ID NO.1-2.

[0038] The upstream primer of the detection primer: SEQ ID NO.1, Sox10-F: 5'-GGAGAGAGAGATGTTAGGTAGGTAG-3'.

[0039] The downstream primer of the detection primer: SEQ ID NO.2, Sox10-R: 5'-AGTTTAATAAAAATTTTCTCTTTCTCTC-3'.

[0040] The product length is 388bp.

[0041] Therefore, the above detection primers based on the methylation of the Sox10 gene in peripheral blood can be applied in the detection of the methylation of the Sox10 gene.

[0042] Of course, the detection primers based on the methylation of the Sox10 gene in peripheral blood can also be applied to the preparation of a detection kit for the methylation of the Sox10 gene.

Embodiment 2

[0044] This embodiment provides a Sox10 gene methylation detection kit, which includes detection primers based on Sox10 gene methylation in peripheral blood. The nucleotide sequences of the detection primers are shown in SEQ ID NO.1-2.

[0045] The upstream primer of the detection primer: SEQ ID NO.1, Sox10-F: 5'-GGAGAGAGAGATGTTAGGTAGGTAG-3'.

[0046] The downstream primer of the detection primer: SEQ ID NO.2, Sox10-R: 5'-AGTTTAATAAAAATTTTCTCTTTCTCTC-3'.

[0047] The product length is 388bp.

[0048] The detection kit includes at least one of genomic DNA extraction reagents, sodium bisulfite, sodium acetate, absolute ethanol, and PCR reaction reagents.

[0049] Genomic DNA extraction reagents include PBS buffer, proteinase K and STE solution.

[0050] The STE solution was prepared from 0.1mol / L NaCl, 10mmol / LTris-HCl, 1mmol / LEDTA.

[0051] PCR reaction reagents include PCR reaction buffer, Taq enzyme and ddH2O; PCR reaction buffer includes 100 mMTris-HCl (pH 8.8 at 25°C), 5...

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Abstract

The invention provides detection primers, kits and applications based on Sox10 gene methylation in peripheral blood, belonging to the field of genetic engineering. The detection primers based on the methylation of the Sox10 gene in peripheral blood provided by the present invention can detect the methylation level of the promoter region of the Sox10 gene in a targeted manner, so the detection primers are applied to the detection of the methylation level of the Sox10 gene in peripheral blood and the preparation of Sox10 gene alpha The methylation detection kit has a good effect; the Sox10 gene methylation detection kit is easy to operate, easy to implement, and has a high medical application prospect.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to detection primers, detection kits and applications based on the methylation of Sox10 gene in peripheral blood. Background technique [0002] The Sox10 gene is located at 22q13.1, the full-length cDNA is 1.4kb, and contains 5 exons, of which only exons 3, 4, and 5 encode proteins, and the 5' non-coding region contains one or more non-coding exons . Sox10 was first expressed in the dorsal neural tube in the early stage of neural crest stem cell migration, and Sox10 began to express at day 9.5 (E9.5) of mouse embryonic development, and its expression began to decrease at day 16.5 (E16.5). Then, as neural crest stem cells gradually differentiate, they are expressed in their derivatives such as peripheral nervous system, gastrointestinal system and melanocytes. Sox10 is widely expressed in various tissues and organs in humans and adult mice, such as brain, heart, lung, adrenal gla...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/154
Inventor 张树成程兰杰黄文凯
Owner SHENGJING HOSPITAL OF CHINA MEDICAL UNIV
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