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Analysis of COL4A3 Gene (Mutation) by RT-qPCR (CAT#: STEM-MT-1528-LGZ)

Introduction

Official Full Name: collagen type IV alpha 3 chain<br />Also known as: ATS2; ATS3; BFH2<br />Type IV collagen is the main structural component of the basement membrane and is a multimeric protein composed of three α subunits. These subunits are encoded by 6 different genes, α1 to α6, and each subunit can form a triple helix structure with the other 2 subunits to form type IV collagen. This gene encodes α3. In Goodpasture syndrome, autoantibodies bind to collagen molecules in the alveolar and glomerular basement membranes. The epitopes eliciting these autoantibodies are predominantly localized in the non-collagenous C-terminal domain of the protein. A specific kinase phosphorylates amino acids in the same C-terminal region, and the expression of this kinase is upregulated during pathogenesis. This gene is also associated with an autosomal recessive form of Alport syndrome. The mutations that cause this syndrome are also located within the exons encoding this C-terminal region. Like other members of the collagen IV gene family, this gene is in a head-to-head conformation with another collagen IV gene, so that each gene pair shares a common promoter.




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