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Breast cancer susceptibility gene BRCA2-K2305N mutant and specific primer thereof

A BRCA2-K2305N, susceptibility gene technology, applied in the field of molecular biology, can solve problems such as loss of protein to repair DNA damage, chromosomal instability, etc., and achieve the effect of convenient and easy diagnosis

Active Publication Date: 2021-06-11
WUXI NO 5 PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The protein expressed after the BRCA2 gene mutation loses the function of repairing DNA damage, resulting in chromosomal instability and promoting tumorigenesis

Method used

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  • Breast cancer susceptibility gene BRCA2-K2305N mutant and specific primer thereof
  • Breast cancer susceptibility gene BRCA2-K2305N mutant and specific primer thereof
  • Breast cancer susceptibility gene BRCA2-K2305N mutant and specific primer thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0036] The preparation of embodiment 1 DNA template

[0037] Use purchased kits to extract genomic DNA from whole blood, and the specific steps are as follows:

[0038] (1) Take a sterile 2.0mL centrifuge tube and add 1mL of cell lysate.

[0039] (2) Gently shake the whole blood sample anticoagulated with EDTA until it is thoroughly mixed; then pipette 500 μL of blood sample into the above-mentioned centrifuge tube containing the cell lysate, and gently pour the centrifuge tube 5-6 times to mix well.

[0040] (3) Incubate at room temperature for 10 minutes (during this period, invert the centrifuge tube 2-3 times to mix well).

[0041] (4) Centrifuge at room temperature for 5 minutes at 12000 rpm.

[0042] (5) Use a pipette to slowly remove the supernatant as much as possible, and be careful not to suck out the white substance at the junction of the two phases.

[0043] (6) Vigorously mix with a vortex shaker (Votex) until the leukocytes are resuspended (10-15 seconds).

...

Embodiment 2

[0054] Example 2 BRCA2 gene g.32344631G>T mutation gene detection scheme:

[0055] Instruments: Veriti 96 PCR instrument, BIO-RAD Gel Doc XR+ gel imager (Bio-Rad, USA), gel electrophoresis (Beijing Liuyi Company).

[0056] Reagents: QIAamp DNA extraction kit (Qiagen, Germany); DNA Isolation Kit extraction kit (PELFREEZ, Beijing); PCR buffer, dNTP, Taq enzyme (ABI, USA); primers were synthesized by Shanghai Sangon Biotechnology Co., Ltd.

[0057](1) Primer design: according to the BRCA2 gene (sequence number: NC_000013.11) recorded in the GenBank of the National Center for Biological Information (NCBI), primers were designed by Oligo 6.0 primer software, and finally a pair of specific oligonucleotide primer sequences were determined, The sequence of the forward primer is shown in SEQID NO:3 in Table 1; the sequence of the reverse primer is shown in SEQID NO:4, and the length of the amplified product fragment is 172bp.

[0058] Table 1. Oligonucleotide primer sequences for rele...

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Abstract

The invention discloses a breast cancer susceptibility gene BRCA2-K2305N mutant and a specific primer thereof, and belongs to the technical field of molecular biology. A primer sequence containing a changed mutant gene is designed by applying a PCR-specific primer technology, and for a specific BRCA2 gene g.32344631G>Tmutation site is subjected to PCR amplification detection. The invention provides a new mutation site of a breast cancer related gene, which can be used for early breast cancer screening.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and particularly relates to a breast cancer susceptibility gene BRCA2g.32344631G>T site mutant and a specific primer thereof. Background technique [0002] In recent years, the morbidity and mortality of cancer in my country have been increasing, and malignant tumors have become a major public health problem in China. According to the results released by the my country Cancer Registry Center in 2016, in 2012, there were 3.586 million new cancer cases in my country, 2.187 million deaths, a crude incidence rate of 265 / 100,000, and a mortality rate of 161.5 / 100,000. In the current situation of malignant tumors, breast cancer ranks first among female malignant tumors, with about 273,000 new cases every year, and the age of onset is showing a trend of younger age. It is the number one killer that seriously threatens women's lives. [0003] Breast cancer is a malignant tumor with a strong...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/156Y02A50/30
Inventor 王学东王玥苹周道平郑国沛唐海林顾娟
Owner WUXI NO 5 PEOPLES HOSPITAL
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