A method for detecting structural variation in target regions of the genome

A technology of regional structure and genome, applied in the field of biological genes, can solve the problem of non-existence, and achieve the effect of high sensitivity and high resolution

Inactive Publication Date: 2011-12-21
SUN YAT SEN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, methods that only detect structural variants in s

Method used

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  • A method for detecting structural variation in target regions of the genome
  • A method for detecting structural variation in target regions of the genome
  • A method for detecting structural variation in target regions of the genome

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Experimental program
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Embodiment 1

[0026] Embodiment 1 is to the detection of human HLA regional genome structure polymorphism (referring to figure 2 )

[0027] 1. Genomic DNA was extracted from the cultured HapMap project Beijing Han B lymphocytes (GM18529).

[0028] 2. According to the GS FLX Titanium 3Kb Span Paired End Library Preparation Method Manual, construct a 3Kb genome paired-end library:

[0029] (1) Fragment the genomic DNA with hydroshear (SC10, Cycle20), and recover 3Kb fragments by gel recovery

[0030] (2) DNA fragment end repair. The reaction system is as follows:

[0031]

[0032] Reaction parameters: 25°C, 20min.

[0033] After the reaction, the reaction product was purified with a QIAquick PCR purification kit and dissolved in 35 μl of elution buffer.

[0034] (3) Connect the circular adapter. The reaction system is as follows:

[0035] repaired fragmented DNA

35

2x Quick Ligase Buffer

50

Circular adapter (20uM)

10

quick ligase

5 ...

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Abstract

The invention provides a method for detecting the structural variation of the genome target region. The present invention skillfully combines double-end library construction technology, chip hybridization technology and high-throughput sequencing technology together, so that the structural variation of any interested target region can be detected with high sensitivity and high resolution.

Description

technical field [0001] The invention relates to the field of biological genes, in particular to a method for detecting structural variation of a genome target region. Background technique [0002] Genome structural variation was first defined as genomic insertions, deletions, inversions, and transpositions larger than 1Kb. With the increasing routine of sequencing the human genome, the definitions of structural variation (SV) and copy number variation (CNV) have also been broadened (eg, to >50 bp). There are two main types of traditional methods for detecting genomic structural variation: [0003] One is the technology based on microarray chip hybridization, such as comparative genomic hybridization (aCGH) and single nucleotide polymorphism chip technology (SNP microarrays). The principle of this method is to hybridize two different samples (control sample and experimental sample) with different fluorescent markers, respectively, with the microarray chip. Differences i...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 徐安龙付永贵贺玲瑜周思思李杰
Owner SUN YAT SEN UNIV
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