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Analysis of TNNT3 Gene (Mutation) by RT-qPCR (CAT#: STEM-MT-2145-LGZ)

Introduction

Official Full Name: troponin T3, fast skeletal type<br />Also known as: TNTF; DA2B2; beta-TnTF<br />The binding of calcium(2+) to the trimeric troponin complex initiates the process of muscle contraction. An increase in Ca(2+) concentration induces a conformational change in the troponin complex and transmits it to the tropomyosin dimer located on the actin filament. The altered conformation allows for increased interactions between the myosin head and actin filaments, ultimately generating muscle contraction. The troponin complex has protein subunits C, I and t. Subunit C binds calcium(2+) and subunit I binds actin and inhibits actin-myosin interactions. Subunit T binds the troponin complex to the tropomyosin complex and is also required for Ca(2+)-mediated activation of actomyosin ATPase activity. There are three distinct troponin T genes that encode tissue-specific T subunit isoforms for fast, slow, and cardiac muscle. This gene encodes the fast skeletal troponin T protein; also known as troponin T type 3. Alternative splicing results in multiple transcript variants encoding additional distinct troponin T type 3 isoforms. A developmentally regulated switch occurs between fetal/neonatal and adult troponin T type 3 isoforms. Other splicing variants have been described, but their biological validity has not been determined. Mutations in this gene may cause distal arthrogryposis multiplex congenita type 2B (DA2B).




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