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Method and device for detecting gene fusion

A gene fusion, to-be-detected technology, applied in the field of biology, can solve the problems of inability to accurately locate fusion breakpoints and fusion forms.

Active Publication Date: 2015-09-09
天津诺禾致源生物信息科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The present invention aims to provide a method and device for detecting gene fusion to solve the technical problem that the method for detecting gene fusion in the prior art cannot accurately locate the fusion breakpoint and the fusion form

Method used

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  • Method and device for detecting gene fusion
  • Method and device for detecting gene fusion

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Embodiment 1

[0044] 1) The sample to be tested is the H2228 ALK fusion cell line, which is quantified by a fluorometer (Qubit), and its concentration is 18.2ng / ul; the reverse transcription conditions are: keep at 42°C for 30min; then raise the temperature to 85°C for 5min ; Finally, keep warm at 10°C, and perform reverse transcription on the RNA to obtain cDNA. The configuration of the reverse transcription system is shown in Table 1.

[0045] Table 1

[0046] Reagent volume reverse transcriptase 1μl Reverse transcription reaction solution 2μl

[0047] RNA 7ul(10ng)

[0048] 2) The cDNA amplicon library contains 7 pairs of primers designed for the fusion of ALK and ROS1 genes. The detailed information is as follows:

[0049]

[0050] The cDNA amplification system configuration for cDNA amplification is shown in Table 2. Perform multiplex PCR amplification to obtain the target fragment. The basic steps are as follows: first, pre-denature a...

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Abstract

The invention discloses a method and a device for detecting gene fusion, wherein the method comprises the following steps: S1, extracting RNA of a to-be-detected sample, and performing reverse transcription on RNA to obtain cDNA; S2, designing primers on two sides of a breaking point of a known fusion gene, and taking cDNA as a template for amplifying to form a sequence with a fusion form, and establishing a sequencing library; S3, sequencing the sequencing library by a high-throughput method to obtain a sequence with a fusion form; S4, detecting the sequence with the fusion form, particularly, S41, using comparison software to compare the sequence with the fusion form, obtained from the step 3, with a corresponding reference sequence with the fusion form; S42, judging whether data obtained from the step S41 meet the analysis requirements or not; S43, detecting the known fusion forms. The method has the advantages of being high in sensitivity and specificity and relatively economic.

Description

technical field [0001] The invention relates to the field of biology, in particular to a method and device for detecting gene fusion. Background technique [0002] Gene mutation refers to the sudden and heritable variation phenomenon (gene mutation) that occurs in genomic DNA molecules. From the molecular level, gene mutation refers to the change of base pair composition or sequence in gene structure. Genes are stable enough to replicate themselves precisely as cells divide, but this stability is relative. Under certain conditions, a gene can also suddenly change from its original form of existence to another new form of existence, that is, at a site, a new gene suddenly appears to replace the original gene. This gene is called a mutant gene. New traits which were never present in the progenitor suddenly appear in the expression of the offspring. [0003] Gene mutation is one of the important factors of biological evolution, so the study of gene mutation has a wide range ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G06F19/22C12M1/34
CPCC12Q1/6869G16B30/00C12Q2535/122
Inventor 蒋智曹志生李宗文张广鑫张兰英孟雪红王玉梅尹静妮谭泽民曹银川吴晓朦潘凯
Owner 天津诺禾致源生物信息科技有限公司
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