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Analysis of MPL Gene (Mutation) by RT-qPCR (CAT#: STEM-MT-0099-LGZ)

Introduction

Also known as: MPLV; TPOR; C-MPL; CD110; THPOR; THCYT2<br />In 1990, an oncogene, v-mpl, was identified from a mouse myeloproliferative leukemia virus capable of immortalizing myeloid hematopoietic cells from different lineages. In 1992, the human homologous gene c-mpl was cloned. Sequence data revealed that c-mpl encodes a protein homologous to members of the hematopoietic receptor superfamily. The presence of c-mpl antisense oligodeoxynucleotides inhibits megakaryocyte colony formation. The ligand for c-mpl, thrombopoietin, was cloned in 1994. Thrombopoietin is a master regulator of megakaryocytopoiesis and platelet formation. The protein CD110 encoded by the c-mpl gene is a 635 amino acid transmembrane domain with two extracellular cytokine receptor domains and two intracellular cytokine receptor box motifs. TPO-R-deficient mice are severely thrombocytopenic, emphasizing the essential role of CD110 and thrombopoietin in megakaryocyte and platelet formation. After binding to thrombopoietin, CD110 is dimerized, and the JAK family of non-receptor tyrosine kinases, as well as the STAT family, MAPK family, adapter protein Shc, and the receptor itself are tyrosine-phosphorylated.




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

Congenital amegakaryocytic thrombocytopenia, Essential thrombocythemia, MPL-Related Disorders, Primary myelofibrosis, Thrombocythemia 2, Primary myelofibrosis, Thrombocytopenia

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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