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Method for detecting expression level of BCL11A gene by using Q-PCR technology and application of method

A gene expression level, Q-PCR technology, applied in the direction of recombinant DNA technology, biochemical equipment and methods, DNA / RNA fragments, etc., can solve the problems of inaccurate detection results and interference

Inactive Publication Date: 2014-11-05
SUZHOU UNIV
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Problems solved by technology

However, this method may be interfered by non-specific double-stranded DNA products, so that accurate detection results cannot be obtained

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  • Method for detecting expression level of BCL11A gene by using Q-PCR technology and application of method
  • Method for detecting expression level of BCL11A gene by using Q-PCR technology and application of method
  • Method for detecting expression level of BCL11A gene by using Q-PCR technology and application of method

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Embodiment Construction

[0031] The present invention will be described in detail below with reference to the accompanying drawings and in combination with embodiments.

[0032] A method for detecting the expression level of BCL11A by Q-PCR:

[0033] 1. Construction of quantitative standard plasmid and template preparation

[0034] Standard plasmid preparation for GAPDH internal reference gene: GAPDH gene was amplified from bone marrow samples, connected to pMD19-T vector, and transformed into Escherichia coli DH5α, positive clones were selected by blue-white screening, and sequenced for identification.

[0035] Standard plasmid preparation for the BCL11A gene: the BCL11A gene was amplified from the bone marrow sample, successfully connected to the expression vector pMIGR-his6, and transformed into Escherichia coli DH5α, positive clones were selected by blue-white screening, and sequenced for identification.

[0036] The two constructed plasmids pMIGR-his-BCL11A were amplified, extracted, and the con...

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Abstract

The invention discloses a method for detecting the expression level of a BCL11A gene by using a Q-PCR technology and an application of the method. Amplification primers and probes of BCL11A and GAPDH are designed, and a Q-PCR amplification system is optimized. According to the invention, the established Q-PCR detection method for detecting the expression level of the BCL11A gene is proved to be capable of precisely detecting the expression level of BCL11A in a sample of a leukemia patient and analyzing the correlation between the expression level of BCL11A and treatment remission rate and the correlation between the expression level of BCL11A and prognosis. The method disclosed by the invention can be used for detecting the expression level of the BCL11A gene and providing the basis for diagnosis, treatment remission and prognostic evaluation for leukemia patients in clinic.

Description

technical field [0001] The invention belongs to the field of molecular biology detection, and relates to a detection method of gene expression, in particular to a Q-PCR detection method of BCL11A expression level and application thereof. Background technique [0002] At present, the widely used method for detecting gene expression is to use SYBR Green dye to detect the relative expression level of genes. Although the experimental design of this method is simple, there is no need to design probes and draw standard curves. However, this method may be interfered by non-specific double-stranded DNA products, so accurate detection results cannot be obtained. [0003] Therefore, in order to more accurately detect the expression level of BCL11A in patient samples and provide a better basis for basic and clinical research, we designed specific fluorescent probes and primers for the BCL11A gene, successfully explored the Q-PCR detection conditions, and The qPCR detection standard ...

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6851C12Q2545/114C12Q2561/101C12Q2561/113
Inventor 尹斌毛政伟郑彦文
Owner SUZHOU UNIV
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