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Kit for detecting liver cancer susceptibility genotyping

A technology for detecting kits and susceptibility genes, which is applied in the field of kits for detecting susceptibility genotyping of liver cancer, and can solve problems such as time-consuming, low throughput, and high cost

Inactive Publication Date: 2010-12-29
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

[0114] Example 1: Primer Synthesis

[0115] The primers mentioned in the summary of the invention were customized from Biological Oligonucleotide Chain Synthesis Company (Shenggong).

Embodiment 2

[0116] Embodiment 2: making gene chip

[0117] (1) Probe preparation

[0118] The probes were uniformly prepared with 3× citrate buffer (SSC) to a final concentration of 1500 nM, and sealed and stored at 4°C. As the probe, the sequence used in the gene chip described in the application number 200910203530.5 can be used.

[0119] (2) slide preparation

[0120] 1. Place the slides vertically on the glass rack, and then put the rack in the glass tank.

[0121] 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 .

[0122] 3. Rinse the soaked foil with deionized water 5 times, and finally immerse in deionized water and shake for 5 minutes.

[0123] 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. ...

Embodiment 3

[0132] Embodiment 3: kit preparation

[0133] (1) Amplification solution preparation

[0134] With 25ml (sterilized double distilled water 18.5ul, 10xPCR buffer 2.5ul) buffer solution, primer (10umol / L, upstream and downstream each 1ul) 2ul, dNTP (10mmol / L, dTTP:Dig-dUTP=10:1), press The aforementioned proportions were prepared as an amplification solution. 10umol / L of primers in the system means that the respective concentrations of all primers included in the summary of the invention reach this level.

[0135] (2) Taq enzyme (polymerase) (2 U / ul) (purchased from TAKARA).

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PUM

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Abstract

The invention provides a kit for liver cancer susceptibility genotyping, which comprises a gene chip, amplification liquid and a polymerase, wherein the amplification liquid consists of multiple primers, the length of each primer is 15-30bp, and the primers are designed according to a base pairing principle and are located at the upstream or downstream of a nucleotide sequence in which all genotype mutation sites locate. The segments in which the mutation sites locate can be amplified through multiple PCR (Polymerase Chain Reaction), which is convenient for detection. The kit overcomes the defects of low efficiency, complicated operation, poor controllability, large genotyping limit and the like in the traditional genotyping method, and a genotyping kit with large flux and high efficiency, prepared by adopting the modern biological gene chip detection technology, can maximally include known susceptible gene sites, and can improve the detection sensitivity through specially-designed primers. The kit for the liver cancer susceptibility genotyping is helpful to carry out rapid genotyping on samples, and plays a great role in the development of evidence-based medicine of liver cancer.

Description

technical field [0001] The invention relates to a nucleic acid typing detection product, in particular to a kit for detecting liver cancer susceptibility genotyping, and discloses its use method. 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 earlie...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
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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