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JAK2 V617F mutation detection application

A V617F, mutation-type technology, applied in the field of detection kits and primer pairs, can solve the problems of being unsuitable for large-scale promotion and use, time-consuming sequencing equipment and high cost of reagents, and achieve low cost, time-saving detection and high efficiency.

Active Publication Date: 2014-11-05
AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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Problems solved by technology

[0003] The current method for detecting this point mutation is mainly sequencing after polymerase chain reaction (PCR), but the sequencing in this method is time-consuming and requires high equipment and reagent costs, which is not suitable for large-scale promotion and use

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  • JAK2 V617F mutation detection application
  • JAK2 V617F mutation detection application
  • JAK2 V617F mutation detection application

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Embodiment Construction

[0025] The present invention will be illustrated through specific examples. Wherein, if there are any unfinished parts, you can refer to experimental manuals such as "Molecular Cloning Experiment Guide (Third Edition)" (Science Press, Beijing) and commercially available Instructions for use of reagents and instruments.

[0026] The inventor commissioned Shanghai Sangon Bioengineering Technology Service Co., Ltd. to synthesize the wild-type JAK2 gene (abbreviated as wild-type) and the JAK2 V617F mutant gene with G 1849T mutation (abbreviated as is the mutant type), and the mutant gene is mixed into the wild-type gene in a certain proportion (0.1%, 1%, 10%) to form different samples to be tested.

[0027] Simultaneously, the inventor commissioned Shanghai Sangon Bioengineering Technology Service Co., Ltd. to synthesize the following primers:

[0028] Primer 1: 5'-TGAAGCAGCAAGTATGATGAG-3';

[0029] Primer 2: 5'-GGGAGTATGTGTCTGTGGAGACTGACACCTAGCTGTGATCCTG-3'.

[0030] Use TaKaRa ...

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Abstract

The invention belongs to the field of nucleic acid detection, and particularly relates to the detection of JAK2 (Janus Kinase 2) gene V617F locus DNA mutation (G1849T) and a test kit and a primer pair used for the detection. The present invention discloses a method for detecting V617F mutation in the JAK2 gene, which comprises (1) performing PCR amplification for the samples to be tested, and (2) performing melting analysis by using a melting analyzer. Further, the present invention also discloses the test kit and the primer pair for the detection method.

Description

technical field [0001] The invention belongs to the field of nucleic acid detection, in particular, the invention relates to the detection of the deoxyribonucleic acid mutation (G1849T) at the V617F site of the JAK2 gene, the detection kit used, the pair of primers and the like. Background technique [0002] Janus kinase 2 (hereinafter referred to as JAK2) belongs to the Janus kinase family and is an intracellular tyrosine protein kinase. From the N-terminal to the C-terminal, JAK2 is composed of seven highly conserved homology domains (JH1~JH7), JH7-JH1, and acts on EPO, TPO, IL-3, G-CSG, and GM-CSF. It plays a role in regulating intracellular signal transduction and producing cell proliferation effects. However, the 1849th guanine in exon 14 of the JAK2 gene was replaced by thymine, which mutated the 617th phenylalanine in the JAK2 protein pseudokinase region to valine (JAK2 V617F mutation), which would cause the loss of the pseudokinase region. Live, eventually lead to ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 关明吴之源张心菊刘维薇陈宇明张伟万峻于波
Owner AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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