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Glioma mutant gene detection primer composition, kit and application of kit

A technique for detecting glioma and primers, which is applied in the field of genetic engineering, can solve problems such as inconvenience, and achieve the effects of cost reduction, convenient operation, and low overall cost

Inactive Publication Date: 2021-08-24
上海贞固医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the differences in the principle, reagents, and sequence and length of Barcode and UID fragments of each sequencer manufacturer, the library construction reagent (hybridization capture probe library / multiple PCR primer pool) used on a sequencing platform is designed. ) are generally not suitable for use on other platforms, causing some inconvenience

Method used

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  • Glioma mutant gene detection primer composition, kit and application of kit
  • Glioma mutant gene detection primer composition, kit and application of kit
  • Glioma mutant gene detection primer composition, kit and application of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The verification method adopted in the multiple PCR library construction and sequencing process of the present invention is as follows:

[0041] 1. Sample Preparation

[0042] 1.1 Sample extraction

[0043] Coriell's commercial reference product NA12878 and clinical samples from the hospital were extracted and quantified by Qubit.

[0044] Purchase the commercial reference product EGFR vIII standard from Kebai Company.

[0045] Cell line 1p19q standards from the laboratory were extracted and quantified by Qubit.

[0046] 1.2 Sample quality control

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PUM

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Abstract

The invention is applicable to the technical field of gene engineering, and provides a glioma mutant gene detection primer composition, a kit and application of the kit. The primer composition comprises 242 pairs of primers or a combination comprising part of the primers, and specific sequences in a forward primer and a reverse primer of each pair of primers are selected from sequences shown as SEQ ID NO.1-484. The specific sequences in the forward primer and the reverse primer of each pair of primers are 18-25 base sequences in the sequence. A forward primer of the amplification primer pair comprises a sequence as shown in SEQ ID NO. 485. A reverse primer of the amplification primer pair comprises a sequence as shown in SEQ ID NO. 486. According to the invention, a plurality of authoritative databases are combined, mutation hot spot regions of 13 glioma related genes and conserved regions of 1p and 19q chromosomes are selected, a low-initial-quantity or low-quality FFPE sample can be detected, a cerebrospinal fluid sample can also be detected, and general requirements of diagnosis, medication and prognosis of glioma are met. And the comparison rate, the target rate and the uniformity of amplicons are relatively good.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a primer composition for detecting glioma mutation genes, a kit and the application of the kit. Background technique [0002] Next-generation sequencing technology (Next-Generation Sequencing, NGS), also known as next-generation sequencing technology, is a high-throughput detection technology that can sequence hundreds of thousands or even hundreds of millions of DNA molecules at a time. It is used in prenatal detection, tumor mutation analysis and medication guidance, pathogenic microorganism screening and other fields. The Illumina platform is now the most commonly used NGS sequencer in China, with a relatively high market share. The Ion Torrent platform and the domestic MGI platform also have their own unique advantages, and the domestic application ratio is also growing steadily. [0003] Before NGS sequencing, it is usually necessary to enrich and a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6806C12Q1/6869C12N15/11
CPCC12Q1/6886C12Q1/6806C12Q1/6869C12Q2600/156
Inventor 宋钢王朝晖王强
Owner 上海贞固医学检验实验室有限公司
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