Ventricular fibrillation detecting genetic differential diagnosis kit

A technology for differential diagnosis and ventricular fibrillation, applied in the field of molecular biology, can solve the problems of low efficiency, high cost of single nucleotide sequencing, Sanger sequencing method cannot meet the requirements of sequencing, etc., to reduce costs, improve detection accuracy and Stability, avoiding uncertainty about the effect of primers

Inactive Publication Date: 2018-07-20
FUWAI HOSPITAL CHINESE ACAD OF MEDICAL SCI & PEKING UNION MEDICAL COLLEGE
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AI Technical Summary

Problems solved by technology

[0005] At present, Sanger sequencing, as a traditional sequencing technology, can only determine one base sequence in one reaction. Although it is accurate, the efficiency is low, and the cost of single nucleotide sequencing is high.
For the research of complex diseases with genetic heterogeneity, it is usually necessary to sequence the entire coding regions of several or even dozens of genes to find valuable mutation information, and Sanger sequencing cannot meet the requirements of sequencing

Method used

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  • Ventricular fibrillation detecting genetic differential diagnosis kit
  • Ventricular fibrillation detecting genetic differential diagnosis kit
  • Ventricular fibrillation detecting genetic differential diagnosis kit

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Embodiment Construction

[0035] 1. The present invention is described in further detail and completeness below by means of the examples, but the present invention is not limited to the scope of the examples.

[0036] 2. Use TIANamp Blood DNA Midi Kit (DP332) to extract the genome of the sample to be tested. The volume of elution buffer (TE) is not less than 200 μL, and the OD260 / 280 value reaches 1.8-2.0. Take 2 μL of 25ng / μL DNA as a starting point. start template.

[0037] 3. Quantify the sample with Qubit, and use ddH after quantification 2 O Dilute the g DNA sample to 25 ng / μL.

[0038] 4. Building a database

[0039] 5. DNA fragmentation of genomic samples and addition of adapters and tags

[0040] 6. Add 1.5ml of Ethanol to 4.5ml of TargetCap Stop Solution, and mix thoroughly to make the final concentration of Ethanol 25%. Bring 1X TargetCap Stop Solution to room temperature.

[0041] 7. Take out the Purification Beads and place them at room temperature for at least 30 minutes.

[0042] 8....

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Abstract

The invention discloses a ventricular fibrillation detecting genetic differential diagnosis kit. The ventricular fibrillation detecting genetic differential diagnosis kit is characterized by being a genetic differential diagnosis kit for detecting ventricular fibrillation by a massively parallel sequencing platform NGS technology. The ventricular fibrillation detecting genetic differential diagnosis kit has the main characteristics as follows: A, capture probes are uniquely designed and prepared for all exon fragments of 91 genes; B, unique connectors with label sequences are designed; C, universal primers perform PCR amplification on probe sequences; D, unique operation of capturing mixed target segments is designed.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medical diagnosis and biotechnology, relates to an in vitro diagnostic kit for ventricular fibrillation gene mutation, and is a kind of ventricular fibrillation gene differential diagnosis kit applied to the NGS technology of a new generation sequencing platform. Background technique [0002] Ventricular fibrillation (VF), referred to as ventricular fibrillation, refers to ventricular muscle fibrillation, the regular and orderly excitation of the ventricle and the disappearance of systolic and systolic functions, resulting in the loss of the heart's blood pumping function, resulting in a sharp drop in blood pressure to zero. Ventricular fibrillation is a functional heart stopped beating. Ventricular fibrillation is one of the most serious complications of acute myocardial infarction. In the acute phase of myocardial infarction, the incidence of ventricular fibrillation is 15%, and abou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/156
Inventor 王继征宋雷惠汝太张禅那邹玉宝
Owner FUWAI HOSPITAL CHINESE ACAD OF MEDICAL SCI & PEKING UNION MEDICAL COLLEGE
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