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Human multi-gene mutation detection kit based on high-throughput sequencing method

A detection kit and the technology of the kit, which are applied in the biological field, can solve the problems of abnormal termination of DNA polymerase, increased economical and time cost, and high reagent cost, and achieve the effects of low detection limit, high sensitivity and high specificity

Inactive Publication Date: 2017-09-15
SINGLERA GENOMICS (SHANGHAI) LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] Disadvantages: The composition or secondary structure of the DNA template sometimes causes abnormal termination of the DNA polymerase
[0019] Disadvantages: small coverage, high reagent cost
The combination of DNA mutation and RNA fusion detection will increase economic and time costs

Method used

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  • Human multi-gene mutation detection kit based on high-throughput sequencing method
  • Human multi-gene mutation detection kit based on high-throughput sequencing method
  • Human multi-gene mutation detection kit based on high-throughput sequencing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Composition and preparation of embodiment 1 kit

[0057] The human multigene mutation detection kit (high-throughput sequencing method) of the present invention includes a DNA PCR primer mixture and an RNA PCR primer mixture, as shown in the following table 1-1, and the DNA PCR primers include KRAS gene mutation detection primers Right, EGFR gene mutation detection primer pair, described RNA PCR primer comprises ALK gene fusion detection primer pair, described KRAS gene mutation detection primer pair comprises forward primer and nucleoside as shown in SEQ ID NO.1 of nucleotide sequence The reverse primer whose acid sequence is shown in SEQ ID NO.2, the EGFR gene mutation detection primer pair includes the first detection primer pair, the second detection primer pair, the third detection primer pair and the fourth detection primer pair, the first detection primer pair A detection primer pair includes a forward primer with a nucleotide sequence as shown in SEQ ID NO.3 and...

Embodiment 2

[0063] Embodiment 2 kit and its use performance evaluation test

[0064] 1. The composition of the kit:

[0065] This embodiment provides a specific implementation of the kit of the present invention. In this specific implementation method, the main components of the human polygenic mutation detection kit (high-throughput sequencing method) of the present invention are as follows Table 1-2 shown, but not limited thereto, as long as the human polygenic mutation detection kit (high-throughput sequencing method) includes the DNAPCR primers and RNA PCR primers shown in Table 1-1:

[0066] Table 1-2

[0067] main components

Specification

Library Primer Mix

192μL / tube, 1 tube

HIFI PCR Mixture

384μL / tube, 1 tube

FuPa reagent

192μL / tube, 1 tube

ligation buffer

384μL / tube, 1 tube

DNA ligase

192μL / tube, 1 tube

HIFI Enzyme Mix

1600μL / tube, 3 tubes

DNA PCR Primer Mix

192μL / tube, 1 tube

RNA P...

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PUM

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Abstract

The invention belongs to the technical field of biology and particularly relates to a human multi-gene mutation detection kit based on a high-throughput sequencing method and an application thereof. The kit includes a KRAS gene mutation detection primer pair, an EGFR gene mutation detection primer pair and an ALK gene fusion detection primer pair. The kit can simultaneously detect DNA mutation and RNA fusion, is high in sensitivity and low in detection limit, is high in specificity, has good repeatability, and can reach 100% in both accuracy and positive coincident rate.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a human multi-gene mutation detection kit (high-throughput sequencing method) and its application. Background technique [0002] Lung cancer is one of the common malignant tumors that seriously endanger human health, 80%-85% of which are non-small cell lung cancer (NSCLC). With the deepening of research on tumor pathogenesis and its biological behavior, targeted therapy has become a research hotspot, including multiple gene targets such as EGFR, KRAS, and ALK. [0003] Epidermal growth factor receptor (EGFR) is an important target for tumor therapy. The EGFR gene is located on the short arm of human chromosome 7 (7p12), with a length of about 118kb, consisting of 28 exons, and the intracellular region of the transcellular membrane glycoprotein transcribed by it has tyrosine kinase (TK) activity , the signal can be transmitted to the nucleus through two pathways, one is ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C40B50/06
CPCC12Q1/686C12Q1/6869C12Q1/6886C12Q2600/156C40B50/06C12Q2535/122C12Q2531/113C12Q2537/143
Inventor 刘蕊
Owner SINGLERA GENOMICS (SHANGHAI) LTD
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