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Analysis of BRCA1 Gene (Mutation) by RT-qPCR (CAT#: STEM-MT-2482-LGZ)

Introduction

Official Full Name: BRCA1 DNA repair associated<br />Also known as: IRIS; PSCP; BRCAI; BRCC1; FANCS; PNCA4; RNF53; BROVCA1; PPP1R53<br />This gene encodes a 190 kD nuclear phosphorylated protein that plays a role in maintaining genome stability and which also acts as a tumor suppressor. The BRCA1 gene contains 22 exons and is approximately 110 kb in length. The encoded protein associates with other tumor suppressors, DNA damage sensors, and signal transduction to form a large multisubunit protein complex known as the brca1-associated genomic surveillance complex (BASC). This gene product associates with RNA polymerase II and interacts with the histone deacetylase complex through the C-terminal domain. Thus, this protein plays a role in transcription, DNA double-strand break repair, and recombination. About 40% of hereditary breast cancers and more than 80% of hereditary breast and ovarian cancers are caused by mutations in this gene. Alternative splicing plays an important role in regulating the subcellular localization and physiological function of this gene. Many alternatively spliced transcript variants, some of which are disease-associated mutations, have been described for this gene, but the full-length nature of only some of these variants has been described. A related pseudogene was also found on chromosome 17.




Principle

Quantitative reverse transcription PCR (RT-qPCR) is an experimental method applied to PCR experiments using RNA as the starting material. In this method, total or messenger RNA (mRNA) is first transcribed into complementary DNA (cDNA) by reverse transcriptase. Subsequently, qPCR reaction was performed using cDNA as template.

Applications

Gene mutation analysis.

Procedure

1. Sample processing and preparation of PCR reaction system.
2. Add the amplification template, cover the PCR reaction cover, mix well, centrifuge at low speed instantaneously, and transfer to the PCR instrument.
3. Set the program for PCR amplification.
4. Data analysis.

Materials

Sample: depends on the customer's analysis requirements