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Microsatellite colorectal cancer instability amplification system and detection kit thereof

An amplification system and unstable technology, applied in the field of PCR amplification system, can solve limited problems, achieve convenient operation, large-scale promotion and application, and intuitive analysis of PCR results

Inactive Publication Date: 2015-02-25
上海赛安生物医药科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are many deficiencies in cost control and operability for multiple fluorescent labels, and the addition of sex-determining sites is also very limited in preventing experimental operators from confusing samples.

Method used

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  • Microsatellite colorectal cancer instability amplification system and detection kit thereof
  • Microsatellite colorectal cancer instability amplification system and detection kit thereof
  • Microsatellite colorectal cancer instability amplification system and detection kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] The colorectal cancer microsatellite instability detection kit of this embodiment is composed of at least six test tubes containing corresponding reagents fixed in a paper packaging box. The six test tubes are respectively an amplification buffer test tube and a Mg 2+ Test tube, dNTP test tube, hot start enzyme test tube, primer mixture test tube and ultrapure water test tube. The order in which the test tubes are arranged in the packaging box is not particularly required, as long as they are clearly marked.

[0049] The development principle and use method of the detection kit of this embodiment are:

[0050] A. Design, synthesis and screening of primers for MSI gene locus:

[0051] According to the Bethesda standard microsatellite locus, including five genes of BAT25, BAT26, D2S123, D5S346, and D17S250, the polymorphic microsatellite marker primers were designed.

[0052] Design primers and use Oligo7.0 software for performance evaluation. Firstly, make the Tm value of each p...

Embodiment 2

[0097] The rest of the colorectal cancer microsatellite instability detection kit of this example is the same as that of Example 1, except that the concentrations of the forward primer and the reverse primer in the universal-specific chimeric primer pair are different.

Embodiment 3

[0099] The rest of the colorectal cancer microsatellite instability detection kit of this example is the same as that of Example 1, except that the concentrations of the forward primer and the reverse primer in the universal-specific chimeric primer pair are different.

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Abstract

The invention relates to a microsatellite colorectal cancer instability amplification system and a detection kit thereof. The amplification system disclosed by the invention comprises a general-specific chimeric primer pair and a mixture of general primers, wherein the general-specific chimeric primer pair respectively aims at microsatellite sites BAT25, BAT26, D2S123, D5S346 and D17S250; the general primer is FluD5-Up; respective positive primer 5' ends in the general-specific chimeric primer pair are provided with a sequence of the FluD5-Up; the FluD5-Up is a fluorescent-labeled primer. The amplification system and the detection kit thereof which are disclosed by the invention overcome the shortcoming that different primers require for multiple fluorescent labels; five microsatellite gene sites are detected in the same PCR (polymerase chain reaction) system, so that a result is intuitive and reliable; the time can be saved, and the efficiency is high; MSI (medium-scale integration) gene sites can be effectively detected and analyzed.

Description

Technical field [0001] The invention relates to a PCR amplification system and a detection product using the amplification system, and belongs to the field of biotechnology. Background technique [0002] The incidence of colorectal cancer is gradually increasing at home and abroad, and it is one of the common malignant tumors that seriously endanger human health. Studies have found that the instability of the genome plays an important role in the pathogenesis of colorectal cancer. Genome instability includes chromosomal instability (CI) and microsatellite instability (MSI). Previous studies on the incidence of colorectal cancer have shown that chromosome instability is the main cause of colorectal cancer, and chromosome instability refers to the increase or decrease in the number of all chromosomes. In recent years, studies have found that microsatellite instability is another important mechanism of colorectal cancer. [0003] Microsatellites, also known as short tandem repeats (...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/156
Inventor 赵新泰王明
Owner 上海赛安生物医药科技股份有限公司
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