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Detection of JAK2 V617F Mutation by RT-qPCR (CAT#: STEM-MT-0024-LGZ)

Introduction

JAK2 V617F mutations have been associated with a number of diseases primarily associated with myeloproliferative disorders, including polycythemia vera, essential thrombocythemia and idiopathic myelofibrosis, increasing the need for detection using molecular techniques. The latter are varied and all aim to simplify the methods used for detection.




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

Myeloproliferative Disorders

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