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Analysis of LAMC2 Gene (Mutation) by RT-qPCR (CAT#: STEM-MT-1097-LGZ)

Introduction

Official Full Name: laminin subunit gamma 2
Also known as: B2T; CSF; EBR2; BM600; EBR2A; JEB3A; JEB3B; LAMB2T; LAMNB2
Laminins are a class of extracellular matrix glycoproteins that are the major non-collagenous components of basement membranes. They are involved in a variety of biological processes including cell adhesion, differentiation, migration, signaling, neurite outgrowth and metastasis. Laminins, composed of 3 non identical chains: laminin alpha, beta and gamma (formerly A, B1, and B2, respectively), have a cruciform structure consisting of 3 short arms, each formed by a different chain, and a long arm composed of all 3 chains. Each laminin chain is a multidomain protein encoded by a different gene. Several isomers of each chain have been described. Different alpha, beta and gamma chain isomers combine to give rise to different heterotrimeric laminin isoforms which are designated by Arabic numerals in the order of their discovery, i.e. alpha1beta1gamma1 heterotrimer is laminin 1. The biological functions of the different chains and trimer molecules are largely unknown, but some chains have been shown to differ in their tissue distribution, likely reflecting distinct functions in vivo. This gene encodes the gamma chain isoform laminin gamma-2. The gamma-2 chain, previously thought to be a truncated version of the beta chain (B2t), is highly homologous to the gamma-1 chain; but lacks domain VI and is shorter in domains V, IV, and III. It is expressed in several fetal tissues, but unlike gamma-1, and is specifically localized to epithelial cells of the skin, lung and kidney. The γ-2 chain, together with the α-3 and β-3 chains, make up laminin 5 (formerly known as kalinin), a component of the anchoring filament that connects epithelial cells to the underlying basement membrane. The epithelium-specific expression of the gamma 2 chain implied its role as an epithelium attachment molecule, and mutations in this gene have been associated with junctional epidermolysis bullosa, a skin disease characterized by blisters due to disruption of the epidermal -dermal junction. Two transcript variants arising from alternative splicing of the 3' exon have been described and encode distinct γ-2 chain alloforms. These two variants are expressed differently in embryonic tissues, however, the biological significance of these two forms is unknown. Transcript variants utilizing alternative polyA_signals have also been found in the literature.




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
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