Primer for detecting MiR-34b/crs4938723 mutant, and application thereof in preparation of gastric cancer screening kit

A technology of crs4938723 and mutants, applied in the preparation of gastric cancer screening kits, the field of primers for detecting MiR-34b/c rs4938723 mutants, can solve the problem of no risk relationship of gastric cancer and the like

Inactive Publication Date: 2014-07-09
JIANGSU PROVINCE HOSPITAL
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no research report on the relationship between miR-34b / c rs4938723 mutation and gastric cancer risk, and there is no report on the application of miR-34b / c rs4938723 gene mutation kit in gastric cancer screening

Method used

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  • Primer for detecting MiR-34b/crs4938723 mutant, and application thereof in preparation of gastric cancer screening kit
  • Primer for detecting MiR-34b/crs4938723 mutant, and application thereof in preparation of gastric cancer screening kit
  • Primer for detecting MiR-34b/crs4938723 mutant, and application thereof in preparation of gastric cancer screening kit

Examples

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Embodiment 1

[0015] Example 1: Take 3 mL of peripheral blood from a subject, and use a whole blood genomic DNA extraction kit (Tiangen, Beijing, China) to extract the subject's genomic DNA. Subsequent PCR amplification technique was used to amplify the fragment of interest in a 20 μL reaction mixture including: 1.625 mM MgCl 2 , 0.14mM dNTPs, 1U Taq enzyme (MBI Fermentas), 2μL×10PCR buffer (MBI Fermentas), 200ng of the subject’s genomic DNA and 10uM primers (sense: 5'-CCTCTGGGAACCTTCTTTGACCAAT-3', antisense: 5' -TGAGATCAAGGCCATACCATTCAAGA-3'). The PCR reaction conditions were initially 95°C denaturation for 5 minutes, 95°C for 30 seconds, 54°C for 30 seconds, 72°C for 1 min, a total of 34 cycles to amplify DNA, and finally 72°C for 10 minutes. A 148bp PCR product of miR-34b / c rs4938723 was obtained. The amplified PCR product was continuously digested with restriction enzyme MLuCI (New England BioLabs, Beverly, MA) at 37°C for 16 hours, then separated on an 8% polyacrylamide gel, and 1.0 ...

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Abstract

The invention discloses a primer for detecting a MiR-34b/crs4938723 mutant, and an application thereof in preparation of a gastric cancer screening kit. The primer comprises a sense primer shown in SEQ ID NO.1 and an antisense primer shown in SEQ ID NO.2. The application value of the MiR-34b/crs4938723 in preparation of the screening kit for detecting people with gastric cancer risks is expanded, and whether the detected person belongs to the people with gastric cancer risks is determined by detecting the MiR-34b/crs4938723 mutant, so as to heighten vigilance of subjects and clinician.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a primer for detecting MiR-34b / c rs4938723 mutant and its application in preparing a gastric cancer screening kit. Background technique [0002] The occurrence of gastric cancer is a complex, multifactorial, multistep process [Parkin DM (2004) International variation. Oncogene23: 6329-6340.; Correa P (1992) Human gastric carcinogenesis: a multistep and multifactorial process--First American Cancer Society Award Lecture on Cancer Epidemiology and Prevention.Cancer Res52:6735-6740.], studies have shown that the interaction between genetic and environmental factors has led to an increase in the risk of gastric cancer [Wu MS (2005) Host-Environment Interactions: Their Impact on Progression from Gastric Inflammation to Carcinogenesis and on Development of New Approaches to Prevent and Treat Gastric Cancer. Cancer Epidemiology Biomarkers & Prevention 14:1878-1882.]. Genetic pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/156C12Q2600/158C12Q2600/178
Inventor 杨力徐泽宽王连生杨超刘东晓马翔唐然张智弘郭人花徐皓张殿彩王悠南
Owner JIANGSU PROVINCE HOSPITAL
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