Analysis of RAF1 Gene (Mutation) by RT-qPCR (CAT#: STEM-MT-1832-LGZ)

Introduction

Official Full Name: Raf-1 proto-oncogene, serine/threonine kinase
Also known as: NS5; CRAF; Raf-1; c-Raf; CMD1NN
This gene is the cellular homologue of the viral raf gene (v-raf). The encoded protein is a MAP kinase kinase kinase (MAP3K), which acts downstream of the directly bound membrane-associated gtpase Ras family. Once activated, cellular RAF1 protein can phosphorylate and activate the dual-specificity protein kinases MEK1 and MEK2, which in turn phosphorylate and activate the serine/threonine-specific protein kinases ERK1 and ERK2. Activated ERKs are pleiotropic factors in cell physiology and play an important role in regulating the expression of genes related to cell division cycle, apoptosis, cell differentiation and cell migration. Mutations in this gene are associated with Noonan syndrome 5 and LEOPARD syndrome 2.

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Principle Applications Procedure Materials Notes Related Products

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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