Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Kit for detecting B-raf gene polymorphism by pyro-sequencing method and method

A pyrosequencing method and gene polymorphism technology, applied in the field of molecular biology, to achieve the effect of small sample size and easy operation

Active Publication Date: 2014-11-19
CENT SOUTH UNIV
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

According to the latest research, K-ras mutation may cause 30%-40% colorectal cancer patients to be ineffective to EGFR-targeted therapy, while B-raf mutation may cause 10%-15% K-raf wild-type patients to develop drug resistance
[0004] Zelboraf, a newly launched drug for the treatment of advanced metastatic or unresectable melanoma, can only be used in patients with B-rafV600E mutation

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
  • Kit for detecting B-raf gene polymorphism by pyro-sequencing method and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] B-raf upstream primer: 5′-CTTTCTAGTAACTCAGCAGCAT-3′ (SEQ ID NO.2);

[0029] B-raf downstream primer: 5′-AGTAAAAATAGGTGATTTTGGTCT-3′ (SEQ ID NO.3);

[0030] Wherein, the 5' of the downstream primer is labeled with biotin;

[0031] B-raf sequencing primer: 5′-CCACTCCATCGAGATT-3′ (SEQ ID NO.4);

[0032] 1. DNA extraction

[0033] 1.1 The preparation and inspection of reagent materials before the experiment are as follows:

[0034] (1) Check the shelf life of the kit and ensure that ethanol has been added to Wash Buffer 1 and 2, and tick the corresponding mark on the bottle; (2) Isopropanol (if not available, it can be replaced by absolute ethanol) and 75% ethanol ; (3) 1.5mL Eppendorf tubes and various pipette tips within the validity period of autoclaving.

[0035] 1.2 Take out the EDTA anticoagulant tube containing whole blood from the refrigerator at 4°C, and mix it upside down several times;

[0036] 1.3 Make a mark on the 1.5mL Eppendorf tube corresponding to the...

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 discloses a kit for detecting B-raf gene polymorphism by a pyro-sequencing method and a method. By the kit, the B-raf gene rs113488022(V600E) single nucleotide polymorphism is detected. The kit comprises primers which are shown as SEQ ID NO.2-4. By the kit, the accurate, quick and high-throughout detection of B-raf gene mutation can be realized, so that the safe, reasonable and effective individual administration of a monoclonal antibody (such as cetuximab) of an anti-epidermal growth factor receptor, melanin inhibitor Zelboraf and the like can be realized.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a kit and a method for detecting B-raf gene polymorphism by pyrosequencing. Background technique [0002] B-raf is an oncogene, which encodes a serine / threonine-specific kinase, an important transduction factor of the RAS / RAF / MEK / ERK / MAPK pathway, and participates in the regulation of various biological reactions in cells, such as Cell growth, differentiation and apoptosis, etc. Studies have shown that in a variety of human malignant tumors, such as malignant melanoma, colorectal cancer, lung cancer, thyroid cancer, ovarian cancer, liver cancer and pancreatic cancer, there are different proportions of B-raf gene mutations, about 66% of malignant melanoma and 15% of colon cancers have somatic missense mutations in the B-raf gene. About 80-90% of B-raf gene mutations occur at 1799 nucleotides of exon15, T is mutated to A, resulting in the substitution of glutamic acid...

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
Inventor 周宏灏
Owner CENT SOUTH UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com