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Method for constructing library for full-length genome sequencing of novel coronavirus SARS-CoV-2

A genome sequencing, coronavirus technology, applied in chemical libraries, biochemical equipment and methods, combinatorial chemistry, etc., can solve the problems of inability to detect and analyze virus mutation, incomplete gene region, poor specificity, etc., to improve connection efficiency, The effect of high sensitivity and high detection rate

Inactive Publication Date: 2020-11-24
上海融享生物科技有限公司
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Problems solved by technology

[0005] The purpose of the present invention is to provide a library construction method for the full-length genome sequencing of the new coronavirus SARS-CoV-2, to establish a library of complete mutation information genomes for gene sequencing, to meet the requirements of NGS technology, some of the existing ones on the market The detection sensitivity and specificity of the new coronavirus nucleic acid sequencing library construction kit are low, there are many false positives, and the detected gene region is not comprehensive, so it is impossible to further detect and analyze the mutation of the virus

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  • Method for constructing library for full-length genome sequencing of novel coronavirus SARS-CoV-2
  • Method for constructing library for full-length genome sequencing of novel coronavirus SARS-CoV-2

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Embodiment Construction

[0051] The present invention will be described in detail below in conjunction with specific embodiments.

[0052] The present invention provides a method for constructing a library for sequencing the full-length genome of novel coronavirus SARS-CoV-2, which specifically includes the following steps:

[0053] S1. cDNA synthesis: extract viral RNA, random primer hexamer, viral RNA, and nuclease-free water to prepare a reaction system, mix them in a PCR tube on ice, and place them in a PCR instrument for the first reaction. After the reaction, the PCR tube Take it out, add Reverse Transcription mix G2 (reverse transcriptase reaction solution), mix well and put it in the PCR instrument again to carry out the reverse transcription product from the secondary reaction according to the set temperature program. The specific operation method is as follows:

[0054] (1) Take 1pg ~ 500ng of the extracted viral RNA and configure the system on ice in a clean PCR tube according to the compos...

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Abstract

The present invention relates to a method for constructing a library for full-length genome sequencing of novel coronavirus SARS-CoV-2. A complete mutation information genome library is established for gene sequencing to meet the requirements of an NGS technology, and solve the problems that some existing novel coronavirus nucleic acid sequencing library conducting kits have low detection, sensitivity poor specificity and more false positives, and the detected gene regions are not comprehensive, and the virus mutation condition cannot be further detected and analyzed.

Description

technical field [0001] The invention belongs to the technical field of gene sequencing, and in particular relates to a library construction method for sequencing the full-length genome of the novel coronavirus SARS-CoV-2. Background technique [0002] The new coronavirus (Corona Virus Disease 2019, SARS-CoV-2) is only a type of virus containing RNA (Ribonucleic Acid) genetic material, and its specific RNA sequence is a marker for identifying the virus. Specific nucleic acid sequence detection by real-time fluorescent quantitative PCR (RT-PCR) and gene sequencing has been used as a standard for the diagnosis of new coronavirus infection. At present, due to the relatively harsh requirements and cumbersome process of the gene sequencing method, the inspection process takes a long time and the cost is relatively high, therefore, the nucleic acid detection for the new coronavirus is mainly based on the RT-PCR method. [0003] High-throughput sequencing technology NGS (next-gener...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6806C12N15/10C40B50/06
CPCC12N15/1096C12Q1/6806C12Q1/701C40B50/06C12Q2521/107C12Q2531/113C12Q2537/143C12Q2525/191Y02A50/30
Inventor 魏子涵王晓娜喻利
Owner 上海融享生物科技有限公司
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