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Capture probe set for dementia-related gene, kit, library construction method and use thereof

A technology for capture probes and library construction, applied in chemical libraries, biochemical equipment and methods, combinatorial chemistry, etc., can solve the problems of unfavorable dementia-related gene sequencing analysis and high cost, achieve sequencing cost reduction, control sequencing cost, reduce required effect

Active Publication Date: 2018-11-06
XIANGYA HOSPITAL CENT SOUTH UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Sequencing technology can help us gain a more comprehensive understanding of dementia-related genes. However, the target region of dementia-related genes has not been sequenced in the existing sequencing technology. Therefore, to detect dementia-related genes, whole genome sequencing or external Exon sequencing, since more than 200 dementia-related genes have been reported so far, the cost of using conventional Sanger sequencing or whole exome sequencing is high, which is not conducive to high-throughput sequencing analysis of dementia-related genes

Method used

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  • Capture probe set for dementia-related gene, kit, library construction method and use thereof
  • Capture probe set for dementia-related gene, kit, library construction method and use thereof
  • Capture probe set for dementia-related gene, kit, library construction method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1 Capture probe set of dementia-related genes

[0065] This embodiment provides a set of capture probes for capturing dementia-related genes. The genes captured by the capture probe set and the chromosomal coordinate information of the genes are shown in the following table:

[0066]

[0067]

[0068]

[0069] The design of the capture probe set complies with the following rules:

[0070] (a) The length of the probe is 121bp;

[0071] (b) probe design to merge adjacent regions;

[0072] (c) The probe design interval extends 20bp from the target interval to the two wings;

[0073] (d) The probe reference is the sense strand of the genome;

[0074] (e) The consensus sequences on both wings of the probe are shown in SEQ ID NO.1 and SEQ ID NO.2;

[0075] Numbering

Sequence (5'-3')

SEQ ID NO.1

ATCGCACCAGCGTGT

SEQ ID NO.2

CACTGCGGCTCCTCA

[0076] (f) Ensure probe specificity by comparing whole genome information in...

Embodiment 2

[0079] Embodiment 2 is used for the kit of library construction

[0080] This embodiment provides a kit for constructing a library of dementia-related genes, which kit includes the following reagents:

[0081] ① The capture probe set of dementia-related genes provided in Example 1, and the probe set is labeled with biotin; and streptavidin-labeled magnetic beads used to capture the hybridization target sequence.

[0082] ② Adapters for multi-sample hybridization capture, the adapters include at least two groups of Adapters1, Adapters2, Adapters3 and Adapters4;

[0083] Adapters1 is formed by annealing Adapters1-1 and Adapters1-2; Adapters1-1 has the sequence shown in SEQ ID NO.5, and Adapters1-2 has the sequence shown in SEQ ID NO.6;

[0084] The linker Adapters2 is formed by annealing Adapters2-1 and Adapters2-2, Adapters2-1 has the sequence shown in SEQ ID NO.7, and Adapters2-2 has the sequence shown in SEQ ID NO.8;

[0085] Linker Adapters3 is formed by annealing Adapters...

Embodiment 3

[0093] Example 3 Adapter Synthesis Captured by Multiple Sample Hybridization

[0094] Dilute DNA oligonucleotide single-stranded Adapters1-1 and Adapters1-2 to 100 μM with ultrapure water, then mix 100 μl of diluted Adapters1-1 and Adapters1-2 in a PCR tube, and then place it on a PCR instrument, Run the following program: 95°C for 5 minutes, then turn off the PCR instrument and allow it to cool down naturally for 1 hour, the product is the hybrid capture adapter, such as figure 1 shown. The synthetic method of linker Adapters2, linker Adapters3 and linker Adapters4 is the same as linker Adapters1, as respectively Figure 2-Figure 4 shown.

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Abstract

The invention provides a capture probe set for a dementia-related gene, a kit, a library construction method and a use thereof and relates to the technical field of gene detection. The capture probe set contains coding regions of 95 genes and can capture aiming at a specific coding region and perform high-depth sequencing so that target gene sequencing is accurate and a sequencing cost is reduced.The capture probe set can efficiently enrich the target genes in construction of a library, reduce the sample size requirement and also better control the sequencing cost.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to a dementia-related gene capture probe set, kit, library construction method and application. Background technique [0002] Dementia refers to a group of clinical syndromes with cognitive impairment as the main clinical manifestation caused by neurodegeneration, cerebrovascular disease, infection, trauma, tumor, nutritional and metabolic disorders, etc., and is more common in the elderly. In addition to various cognitive impairments such as orientation, memory, learning, language comprehension, and thinking, most patients also exhibit abnormal behavior, which will eventually lead to a decline or loss of their work and social life functions. Dementia has a high prevalence rate, high disability and mortality rate, and is an aging disease. It has become the fourth cause of death in the elderly population in western developed countries after cancer, heart disease and stroke. T...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/11C40B50/06
CPCC12Q1/6883C40B50/06C12Q2600/156
Inventor 沈璐焦彬李鹏飞范嘉庚廖鑫鑫王伟李佳龙
Owner XIANGYA HOSPITAL CENT SOUTH UNIV
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