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Method for detecting copy number variation based on PCR-LDR technology

A copy number variation, one-to-one technology, applied in the biological field, can solve problems such as the difficulty of designing the length of multiple competitive PCR products, and achieve the effects of short experimental period, shortened experimental period and wide applicability.

Inactive Publication Date: 2013-03-20
上海翼和应用生物技术有限公司
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Problems solved by technology

[0007] The technical problem to be solved by the present invention is to overcome the difficulties in the length design of some multiple competitive PCR products in the prior art, and provide a CNVs detection method with low cost, high throughput, low sample demand, high detection sensitivity and wide popularity

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  • Method for detecting copy number variation based on PCR-LDR technology
  • Method for detecting copy number variation based on PCR-LDR technology
  • Method for detecting copy number variation based on PCR-LDR technology

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Embodiment 1

[0040] Using the method described in the present invention, the copy numbers of the VIPR2 gene segment (GeneBank sequence number NT_007741) and reference segments on chromosome 10 and chromosome 5 (GeneBank sequence numbers NT_030059, NT_006576) were detected, wherein the gene of VIPR2 Three PCR detection segments were designed in the segment.

[0041] 1. Design of multiplex PCR primers:

[0042] Design of multiplex PCR primers: We designed 5 pairs of specific primers according to the selection of the test segment and the reference segment, requiring the TM value to be ≥ 62°C and the length of the amplified product to be between 100 and 500 bp. All primers are analyzed by Blast and Oligo mask software to reduce non-specific amplification and primer dimers.

[0043] The sequences of multiple competitive PCR primers in the sequence are underlined, and the bases of the junction sites are indicated in italic lowercase characters.

[0044] The designed nucleic acid sequence is as...

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Abstract

The invention relates to a method for detecting the copy number variation based on multiple competitive PCR of universal fluorescence primers. The method comprises the following steps: 1, providing multiple competitive PCR primers composed of at least one pair of primers in a section to be detected and at least one pair of primers in a reference section; 2, providing a competitive internal control template, wherein there is only one base replacement between the sequence of the internal control template and an actual sequence; 3, carrying out a multiple competitive PCR reaction; 4, carrying out an LDR reaction; and 5, carrying out data analysis. The invention also provides a kit based on the detection method. The kit is suitable for detecting the copy number variation of 5-25 genes / reactions, and is a copy number variation detection scheme having the advantages of rapidness, medium flux and economy.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for detecting copy number variation based on multiple competitive PCR and LDR technology, which is applied to biological science research and clinical molecular diagnosis. Background technique [0002] Copy number variation (copy number variation, CNVs) is a new form of genomic diversity discovered in recent years, which refers to the structural variation of 1kb~3Mb DNA fragments in the genome compared with the reference sequence. CNVs widely exist in the genome, are associated with the occurrence of some genetic diseases, and have an important impact on the variation of human disease resistance and susceptibility phenotypes. This requires the establishment of a fast, accurate and low-cost CNVs typing detection technology. [0003] With the development of biotechnology, different research groups have successively developed many detection methods, mainly including hybridizat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 陆炯肖君华陈轶群
Owner 上海翼和应用生物技术有限公司