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Fluorescence probe for rapid detection of lung cancer ALK gene rearrangement and preparation method

A fluorescent probe and gene rearrangement technology, applied in DNA preparation, biochemical equipment and methods, recombinant DNA technology, etc., can solve problems affecting diagnostic efficiency, long hybridization reagents, etc., and achieve simple formula operation, specificity and sensitivity High reliability and time-saving effect

Inactive Publication Date: 2016-03-02
河南赛诺特生物技术有限公司
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  • Abstract
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  • Claims
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Problems solved by technology

[0004] At present, there are detection kits for lung cancer ALK gene rearrangement on the market, but the hybridization reagent is too long, usually more than 16 hours, which affects the diagnostic efficiency

Method used

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  • Fluorescence probe for rapid detection of lung cancer ALK gene rearrangement and preparation method
  • Fluorescence probe for rapid detection of lung cancer ALK gene rearrangement and preparation method
  • Fluorescence probe for rapid detection of lung cancer ALK gene rearrangement and preparation method

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Embodiment Construction

[0028] The present invention will be further described below in conjunction with specific examples, and these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The test method that does not indicate specific conditions in the following examples, usually according to conventional conditions, the operation steps are as follows:

[0029] 1. Acquisition of probe DNA

[0030] 1. Streak inoculation: Streak inoculate the BAC strain carrying the ALK gene on an agar plate containing antibiotics (chloramphenicol), and culture at 37°C overnight (about 14 hours);

[0031] 2. Initial culture: the next day, pick a monoclonal strain with good growth status and inoculate it in 2ml of LB liquid medium containing antibiotics (chloramphenicol), at a temperature of 37°C, with a shaking speed of about 200rpm, and cultivate for 6-8 hours ;

[0032] 3. Overnight culture: Transfer each 1ml of bacterial liquid to 200ml of cul...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to the technical field of rapid detection of lung cancer related genes. An obtained fluorescence probe for rapid detection of lung cancer ALK gene rearrangement does not contain repeated sequences, can avoid cross reactions and has the advantages of being high in accuracy, good in specificity and low in false positivity, a rapid fluorescence in situ hybridization is utilized for shortening the hybridization time which is originally 16 h to be 2 h, the diagnosis efficiency is greatly improved, and time is saved.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to the technical field of rapid detection of lung cancer-related genes. Background technique [0002] With the development of modern medicine and the renewal of treatment concepts, the treatment of tumors has gradually changed from traditional surgery, radiotherapy and chemotherapy to personalized molecular targeted therapy, and finding a specific and effective therapeutic target is the current research on tumor treatment. one of the hotspots. The fusion gene of Echinoderm microtubule associated protein-like4 (EML4) and Anaplastic Lymphoma Kinase (ALK) is a non-small cell lung cancer (NSCLC) relay epidermal growth factor receptor (EGFR) mutation, Kras (ras One) the molecular targeting site of another specific tumor driver gene after mutation in the gene family. Both the EML4 and ALK genes are located on the short arm of chromosome 2, respectively located in the P21 band and the P23 ba...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C12N15/10
CPCC12Q1/6886C12Q1/6806C12Q1/6841C12Q2600/156C12Q2527/125C12Q2521/101C12Q2521/345C12Q2543/10
Inventor 齐华李三华刘玲玲牛银银孙淼淼徐玉茵梁永波周静
Owner 河南赛诺特生物技术有限公司
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