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Analysis of SPAST Gene (Mutation) by RT-qPCR (CAT#: STEM-MT-1785-LGZ)

Introduction

Official Full Name: spastin<br />Also known as: FSP2; SPG4; ADPSP<br />This gene encodes a member of the AAA (atpase associated with various cellular activities) protein family. Members of this protein family share an ATPase domain and function in a variety of cellular processes, including membrane trafficking, intracellular motility, organelle biogenesis, protein folding, and proteolysis. The use of alternative translation initiation sites in this gene results in a single transcript variant that can generate isoforms with different N-terminal lengths and thus different export efficiencies from the nucleus to the cytoplasm. Furthermore, alternative splicing produces multiple transcript variants that encode isoforms that also differ in other protein regions. An isoform of this gene has been shown to be a microtubule-severing enzyme that regulates microtubule abundance, fluidity, and plus-end distribution. Mutations in this gene cause the most frequent form of autosomal dominant spastic paraplegia 4.




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