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Gene chip for detecting liver cancer susceptible gene typing

A technology for detecting gene chips and susceptibility genes, applied in the field of gene chips for detecting liver cancer susceptibility genotyping, can solve the problems of high cost, low throughput, and time-consuming

Inactive Publication Date: 2009-10-28
道可名康医学发展(上海)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

With the completion of the Human Genome Project, this technology is becoming more and more exhausted in the typing of liver cancer, because liver cancer is a polygenic disease involving many susceptibility genes, and the throughput of this technology is low. Time consuming and expensive

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0159] Embodiment 1: synthetic probe

[0160] Customize the probes mentioned in the Summary of the Invention from a biological oligonucleotide chain synthesis company (such as Sangon).

Embodiment 2

[0161] Embodiment 2: making gene chip

[0162] (1) Probe preparation

[0163] The probes were uniformly prepared with 3× citrate buffer (SSC) to a final concentration of 1500 nM, and sealed and stored at 4°C.

[0164] (2) slide preparation

[0165] 1. Place the slides vertically on the glass rack, and then put the rack in the glass tank.

[0166] 2. Dissolve 50 grams of NaOH in 200ml of deionized water, add 300ml of 95% ethanol (do not use 100% ethanol), prepare 500ml of alkaline washing solution, and completely immerse the slide in the washing solution for at least 2 hours .

[0167] 3. Rinse the soaked foil with deionized water 5 times, and finally immerse in deionized water and shake for 5 minutes.

[0168] 4. Prepare 350 ml of modification solution with 35 ml of poly-L-lysine (poly-L-lysine), 35 ml of sterilized filtered PBS and 280 ml of deionized water.

[0169] 5. After washing the slides in the third step, quickly put them into the modification solution, shake gen...

Embodiment 3

[0177] Embodiment 3: hybridization reaction and detection result

[0178] (1) spot hybridization

[0179] A hybridization solution was prepared, including 0.02% SDS, 5×SSC, 50% deionized formamide, 0.1% N-laurysarosine, 50 mmol·L-1 sodium phosphate, pH 7.0, and 2% blocking agent.

[0180] Use a pipette gun to pipette 2ul Dig-labeled sample PCR amplification products into 10ul hybridization solution, pre-denature at 95°C for 2 minutes, and then cool on ice for 5 minutes. Place the mixed solution to be hybridized on the probe area on the chip, cover it with a cover glass (to prevent water from evaporating during hybridization), and place it in a hybridization oven at 42°C for 1.5 hours.

[0181] After hybridization, take out the slides carefully and immerse them in the slide tank filled with washing solution 1 (2×SSC, 0.1% SDS). Shake 2-3 times, transfer to the slide tank filled with washing solution 2 (1×SSC), shake the slide tank gently for 2-3 minutes, then move the slide i...

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PUM

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Abstract

The invention provides a gene chip for the liver cancer susceptible gene typing. The gene chip is produced by designing a probe of the susceptible gene relative to the liver cancer. The invention adopts the modern biological gene chip technology to prepare the typing chip which has high flux, high efficiency and includes the known susceptible gene sites and improves the detection sensitivity. The liver cancer susceptible gene typing chip can help people to type the gene of the sample and plays an important role in developing the evidence-based medicine of the liver cancer.

Description

technical field [0001] The invention relates to a nucleic acid typing detection product, in particular to a gene chip for detecting liver cancer susceptibility genotyping. Background technique [0002] Liver cancer is one of the common malignant tumors in my country, with a high mortality rate, ranking third in the ranking of malignant tumor deaths after stomach and esophagus; in rural areas in some areas, it occupies the second place, second only to gastric cancer. About 110,000 people die from liver cancer every year in my country, accounting for 45% of the world's liver cancer deaths. [0003] The occurrence of liver cancer is closely related to genes, such as cysteine ​​9, parking protein 2, etc. These gene changes will change the risk of individual liver cancer. High morbidity, high mortality, and the relationship between genes have given rise to a huge impetus for research on the typing of liver cancer susceptibility genes. [0004] The earliest commonly used typing ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C40B40/06
Inventor 穆海东汪宁梅穆宇豪黎飒顾杰锋
Owner 道可名康医学发展(上海)有限公司
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