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Multiple PCR reagent kit detecting breast cancer susceptibility gene mutation and preparation method thereof

A kit and breast cancer technology, which is applied in the field of medical molecular biology, can solve the problem of high prevalence of breast cancer and achieve the effects of avoiding complicated procedures, strong sensitivity and high accuracy

Active Publication Date: 2008-06-18
BEIJING MOKOBIO LIFE SCI CO LTD
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Problems solved by technology

[0006] The BRCA1 gene (breast and ovarian cancer susceptibility gene) was located at 17q21 in 1990, was successfully cloned in 1994, and was confirmed as a susceptibility gene for breast cancer and ovary. The length is 7.8kb, and the gene product is a protein with 1863 amino acids and a zinc finger structure. The BRCA2 gene is located at 13q12-13, consists of 10254 nucleotides, contains 26 exons, and half of the coding sequence is contained in the 11th exon The exon, which encodes a protein containing 3418 amino acids, Gayther et al. found that family members with mutations in the 3.3kb region of exon 11 of the BRCA2 gene have a high prevalence of breast cancer. There are some similarities in structure, but there is currently no evidence that the two have homology

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  • Multiple PCR reagent kit detecting breast cancer susceptibility gene mutation and preparation method thereof

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Embodiment 1

[0028] Embodiment 1, preparation and assembly of the multiplex PCR kit of the present invention

[0029] 1. Design and preparation of primer pair 1 and primer pair 2

[0030] Primer pair 1: BRCA1 F: 5-TAC CCAGTT GGT TGATTT CC-3 (SEQ ID NO: 1)

[0031] R: 5-CTC ACACAG GGGATC AGC ATT C-3 (SEQ ID NO: 2)

[0032] Primer pair 2: BRCA2 F: 5-GGG AAG CTT CAT AAG TCA GTC-3 (SEQ ID NO: 3)

[0033] R: 5-TTT GTAATG AAG CAT CTG ATA CC-3 (SEQ ID NO: 4)

[0034] It was synthesized by an automatic DNA synthesizer and diluted to 50 pmol / μl.

[0035] 2. Assembly of kit

[0036] 10×PCR reaction buffer 0.002-10ml, dNTPs 0.025-10ml, 25mmol / L MgCl 2 0.002ml-10ml; heat-resistant DNA polymerase 0.3μL-1500μL; primer pair 0.5μL-1500μL; primer pair 20.5μL-1500μL, double distilled water 20μL-100mL; BRCA1 exon 11 positive control 15μl-5mL, BRCA2 first Exon 11 positive control 15μl-5mL, BRCA1 exon 11 negative control 15μl-5mL, BRCA2 exon 11 negative control 15μl-5mL.

[0037] Among t...

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Abstract

The invention discloses a multiple PCR kit for detecting the eleventh exon mutation of breast cancer susceptibility gene and a preparation method thereof. The PCR amplification primers of the kit are composed of primers pair 1 which consist of SEQIDNO:1 and SEQIDNO:2 and primers pair 2 which consist of SEQIDNO:3 and SEQIDNO:4. The multiple PCR kit of the invention combines the SSCP technology with the DNA silver staining and can simultaneously screen the mutation conditions of the eleventh exon of BRCA1 gene and the eleventh exon of BRCA2 gene. The invention has the advantages of strong sensitivity, high accuracy, simple and convenient property, etc. and has important significance on the early detection and the forecast of the breast cancer.

Description

technical field [0001] The present invention relates to a kit, in particular to a multiplex PCR kit for detecting the mutation of the 11th exon of breast cancer susceptibility genes (BRCA1 and BRCA2), and the present invention also relates to the preparation method and detection method of the multiplex PCR kit The method belongs to the field of medical molecular biology. Background technique [0002] Breast cancer is one of the most common malignant tumors in women, and its mortality rate is second only to lung cancer, and its incidence rate is on the rise. In my country's coastal economically developed areas, especially in parts of the three major cities of Beijing, Tianjin, and Shanghai, the incidence of breast cancer has ranked first in female malignant tumors; among women in economically developed areas of Western Europe and North America, breast cancer is a major disease. cause of death. [0003] The natural course of breast cancer is the longest in the pre-clinical pe...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 金鑫戴路
Owner BEIJING MOKOBIO LIFE SCI CO LTD
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