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The BRCA1 rs28897672 – Cys61Gly (qPCR) product is used for genotyping the rs28897672 polymorphism located within the human BRCA1 gene encoding a protein involved in DNA repair and classified as a tumor suppressor (Breast cancer type 1 susceptibility protein). The material for analysis are DNA preparations containing human genetic material.
Kit size: 100 reactions
Reaction: allelic discrimination using Taqman probes (FAM: wild-type allele, HEX: allele carrying the Cys61Gly mutation)
Determination: qualitative/quantitative
Tested genome: human (Homo sapiens)
Kit components:
The BRCA1 rs28897672 – Cys61Gly (qPCR) product is based on the Taqman probe genotyping technique. The DNA fragment surrounding the rs28897672 polymorphism is amplified by a pair of primers and detected by a pair of Taqman probes. The FAM-labeled probe is complementary to the DNA region containing adenine (wild-type allele, allele A), while the HEX-labeled probe is complementary to the DNA region containing cytosine (mutant allele, allele C).
During the reaction, these probes compete with each other to bind to the template. The probe that is 100% homologous to the DNA strand binds preferentially to it over the alternative probe. As the reaction progresses, a strong signal appears for the fully homologous probe and a weak signal for the alternative probe. The genotype of the sample is determined by comparing the signal intensity ratio between the two channels (FAM and HEX). For homozygous samples, a strong signal is obtained on one channel and a weak signal on the other channel. For heterozygous samples, which contain binding sites for each probe, an intermediate signal is obtained on both channels.
According to the dbSNP (NCBI) database, the Cys61Gly mutation is currently described under number rs28897672 as an A→C transversion at position 43106487 on chromosome 17. The different nomenclature of the rs28897672 (A→C transversion) compared to the nomenclature of mutations within the gene (T→G transversion) is due to the fact that the BRCA1 gene is located on the chromosome in the opposite orientation to the chromosome numbering. The wild-type allele has adenine (A) at the polymorphic site. The mutated allele has cytosine (C). At the DNA level, the mutation involves the replacement of thymine with guanine, resulting in the replacement of the amino acid cysteine with glycine at position 61 of the BRCA1 protein. Other alternative names for the Cys61Gly mutation are c.181T>G, 300T>G, and C61G.
The Cys61Gly mutation is a missense mutation occurring within the region encoding the RING-type zinc finger motif. The RING motif contains eight conserved amino acids in the form of cysteine and histidine, forming two binding sites for Zn2+ zinc ions. The Cys61Gly mutation removes one of these sites and thus disrupts the formation of the BRCA1 protein homodimer. In the Polish population, the Cys61Gly mutation accounts for approximately 20% of all mutations observed in the BRCA1 gene.
