Unlock instant, AI-driven research and patent intelligence for your innovation.

Application of reagents for detecting mir-129-2 methylation in the preparation of kits for detecting breast cancer

A methylation and kit technology, applied in the field of molecular diagnosis, can solve problems such as insufficient specificity of ordinary PCR

Active Publication Date: 2016-06-01
济南金域医学检验中心有限公司 +1
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, in view of the lack of specificity of ordinary PCR, the applicant provides a non-diagnostic detection method of miR-129-2 methylation using touch-downPCR method for laboratory research on the molecular mechanism of cancer occurrence

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application of reagents for detecting mir-129-2 methylation in the preparation of kits for detecting breast cancer
  • Application of reagents for detecting mir-129-2 methylation in the preparation of kits for detecting breast cancer
  • Application of reagents for detecting mir-129-2 methylation in the preparation of kits for detecting breast cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0015] experiment material:

[0016] Breast cancer cell lines MDA-MB-425, MDA-MB-468, and SK-BR-3 were purchased from the Cell Bank of the Type Culture Collection Committee of the Chinese Academy of Sciences;

[0017] 6 normal human tissue samples, provided by the cooperative hospital;

[0018] Normal adult chromosomal DNA and methylated DNA as controls were purchased from USBiological;

[0019] Primers were synthesized by Shanghai Introvigen;

[0020] All other materials used in the experiment were commercially available.

[0021] Experimental procedure:

[0022] Collect the cultured MDA-MB-425, MDA-MB-468, SK-BR-3 cell lines and extract DNA;

[0023] DNA extraction from plasma samples;

[0024] Unmethylated cytosine in DNA samples was converted to uracil using EpiTect sodium bisulfite kit (Qiagen, Hilden, Germany);

[0025] Using methylated primers

[0026] Forward primer: GAGTTGGGGGATCGCGGACGGT (SEQ ID NO: 1)

[0027] Reverse primer: ATATACCGACTTCTTCGATTCGCCG (SEQ I...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention belongs to the field of molecular diagnosis, and provides an application of a reagent for detecting methylation of miR-129-2 in preparing a kit for detecting a breast cancer. Specifically, the invention provides an application of a primer with a sequence of SEQ ID NO:1-4 in preparing the kit for detecting the breast cancer. Besides, the invention provides a method for detecting the methylation of miR-129-2 by using Touchdown-PCR non-diagnosis.

Description

technical field [0001] The invention belongs to the technical field of molecular diagnosis, and specifically uses primers to detect the methylation of microRNA miR-129-2 to diagnose breast cancer. Background technique [0002] MicroRNA (microRNA) is a class of highly conserved non-coding small RNA molecules, which regulate gene expression by degrading or inhibiting target messenger RNA (mRNA) at the post-transcriptional level. In the human body, the microRNA miR-129 is transcribed into miR-129-1 and miR-129-2, which are located on chromosome 7q32 and 11p11, respectively. It has been reported that the expression of miR-129-2 is down-regulated in various cancers. It has a tumor suppressive effect. [0003] Methylation is an epigenetic modification of genes. Almost all DNA methylation in mammals occurs at the 5' cytosine of CpG dinucleotide sequences. Methylation of CpG island regions is associated with gene transcriptional repression. Gene promoter methylation is considere...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q1/6886C12Q2600/154C12Q2600/178C12Q2525/207
Inventor 汤郡肖莹莹李晓霞
Owner 济南金域医学检验中心有限公司