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Detection method for detecting difference of nucleic acid fragments

A nucleic acid fragment and detection method technology is applied in the field of detection of differences between deoxyribonucleic acid and ribonucleic acid fragments, and can solve problems such as cumbersome detection methods for nucleic acid fragment differences.

Inactive Publication Date: 2016-01-13
上海中优医学检验所有限公司
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  • Abstract
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  • Application Information

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Problems solved by technology

[0006] The purpose of the present invention is to provide a detection method and detection technology with relatively simple operation method, controllable sample contamination and high detection efficiency in view of the cumbersome problem of the above-mentioned nucleic acid fragment difference detection method

Method used

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  • Detection method for detecting difference of nucleic acid fragments
  • Detection method for detecting difference of nucleic acid fragments

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Experimental program
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Embodiment

[0025] The detection method of the nucleic acid fragment difference of embodiment CYP2C19 gene, comprises the following steps:

[0026] 1. Design primers

[0027] A set of primers were designed for the nucleic acid differential region of the CYP2C19 gene for subsequent amplification and extension detection, namely P1, P2, P3, P4, P5 and P6.

[0028] The concrete nucleotide sequence of above-mentioned primer is:

[0029] P1: 5'-TATCAAAATTTAATGTTGGTCTCG-3'24bp

[0030] P2: 5'-ACTAATAAAGGGCCTTGGGTATTG-3'24bp

[0031] P3: 5'-GAGCCCTGAAATAACTAACGAAG-3'23bp

[0032] P4: 5'-ATTGCTTCCTGATCAAAATGGTA-3'23bp

[0033] P5: 5'-TTGATTATTTCCCGGAACCCATA-3'23bp

[0034] P6: 5'-AATTTCCCCATCAAGATATACAA-3'23bp

[0035] 2. Extraction

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Abstract

The invention discloses a detection method for detecting difference of nucleic acid fragments and belongs to the technical field of nucleic acid determination. According to the invention, a designing scheme of a related primer set and a probe for difference of the nucleic acid fragments is provided; a to-be-detected sample nucleic acid is taken as a template; the primer set is applied for amplified reaction for 80 minutes under constant-temperature reaction conditions of 55-90 DEG C; then, visual judgment can be performed according to turbidity or color changing by utilizing fluorescent dye for dyeing, thereby identifying the difference of the nucleic acid fragments. The detection method is simple and convenient to operate, quick in detection and capable of completing all operations including detection without expensive nucleic acid amplifying instruments; and the detection efficiency is improved.

Description

technical field [0001] The invention relates to the technical field of nucleic acid determination, in particular to a detection method for detecting the difference between deoxyribonucleic acid and ribonucleic acid fragments, including nucleic acid fragment detection methods such as single nucleotide polymorphism, nucleic acid fragment repeat number variation, nucleic acid fragment repetition and deletion, etc. . Background technique [0002] The detection of differences in nucleic acid fragments plays a pivotal role in the fields of genetic diagnosis of animal and plant diseases, food analysis, and environmental monitoring. Differences in nucleic acid fragments include, for example, single nucleotide polymorphisms, variations in the repeat number of nucleic acid fragments, duplications and deletions of nucleic acid fragments, abnormal chromosome numbers, and gene methylation. Among them, single nucleotide polymorphism, variation in the number of repeats of nucleic acid fra...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6858
Inventor 傅咏南何骏奇毛丹丹张奕王校邹创
Owner 上海中优医学检验所有限公司