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Kit and probe for detecting lung cancer TUBB3 gene

A technology of detection kits and gene probes, which is used in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve problems such as inaccurate results, and achieve simple operation, high specificity and sensitivity, Well-designed effects

Inactive Publication Date: 2015-01-14
天津蒙特立医疗科技有限责任公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, the detection of TUBB3 gene amplification on the market mainly uses the PCR method. Since the detection is a mixture of tumor and normal cells, the results are not accurate enough, and a more accurate detection method is urgently needed in the market.

Method used

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  • Kit and probe for detecting lung cancer TUBB3 gene
  • Kit and probe for detecting lung cancer TUBB3 gene

Examples

Experimental program
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Effect test

Embodiment 1

[0018] 1. Acquisition of probe DNA

[0019] 1) Streak inoculation: Streak inoculate the BAC strain carrying the TUBB3 gene and the centriole of chromosome 16 on an agar plate containing antibiotics (chloramphenicol), and culture at 37° C. overnight (about 14 hours).

[0020] 2) Initial culture: Pick a monoclonal strain with good growth status the next day, inoculate it in 2ml of LB liquid medium containing antibiotics (chloramphenicol), put it in a shaker at 37°C, set the shaker speed to about 200rpm, and culture 6-8 hours.

[0021] 3) Overnight culture: transfer each 1ml of bacterial liquid to 200ml culture system, place on a shaker at 37°C with a rotation speed of 200rpm, and culture overnight (about 14-16 hours).

[0022] 4) On the next day, a large amount of DNA was extracted and purified according to the operating instructions of the QIAGEN plasmid mass extraction kit.

[0023] 2. Probe labeling

[0024] 1. Digestion of DNA:

[0025] 1) Digest the target DNA according...

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Abstract

The invention relates to kit and probe for detecting a lung cancer TUBB3 gene. The probe includes a group of FISH probes of two colors, wherein the dual-color FISH probe (1) of the group of FISH probes of two colors comprises a red TUBB3 gene probe (2) and a green No.16 chromosome centriole probe; and TUBB3 is Entrez Gene with the ID of 10381. The method for FISH detection on of lung cancer TUBB3 gene amplification comprises the following steps of: conventionally pretreating a lung cancer tissue paraffin section; then carrying out in situ hybridization by adopting the group of probes of two colors, and judging whether the amplification state of a lung cancer-related gene exists or not according to a detected fluorescence signal. The method disclosed by the invention has the advantages of comprehensiveness in design, easiness in operation, high specificity and sensibility and can be used for fast, accurately and intuitively detecting the amplification state of the lung cancer-related TUBB3 gene.

Description

technical field [0001] The invention relates to the technical field of disease-related gene detection, in particular to a FISH detection method for lung cancer TUBB3 gene amplification, related detection probes and a kit. Background technique [0002] Lung cancer is one of the malignant tumors with the fastest increasing morbidity and mortality and the greatest threat to the health and life of the population. In the past 50 years, many countries have reported that the morbidity and mortality of lung cancer have increased significantly. The incidence and mortality of lung cancer in men rank first among all malignant tumors, and the incidence and mortality of lung cancer occupy the second place in women. The etiology of lung cancer is still not completely clear. A large number of data show that long-term heavy smoking has a very close relationship with the occurrence of lung cancer. [0003] According to the growth, invasion and metastasis speed and scope of lung cancer and t...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6841
Inventor 缪为民
Owner 天津蒙特立医疗科技有限责任公司
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