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Detection probe, kit and method for Dystrophin gene

A probe and gene technology, applied in the field of Dystrophin gene detection probes, can solve the problems of easy failure, multiple amplification, PCR failure, etc.

Pending Publication Date: 2020-09-01
BEIJING GRANDOMICS BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the length of the Dystrophin gene is about 2.3Mb, the longest length of PCR amplification is about 10K. If PCR is used, about 230 pairs of PCR primers are needed, and amplification with too many primers is likely to fail.
In addition, if the PCR primers are located on the missing segment, the PCR will fail
Therefore Dystrophin gene is not suitable for PCR amplification method

Method used

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  • Detection probe, kit and method for Dystrophin gene
  • Detection probe, kit and method for Dystrophin gene
  • Detection probe, kit and method for Dystrophin gene

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Embodiment

[0096] 1. Preparation of probes for DMD gene hybridization capture

[0097] For the full-length sequence of the DMD gene, one probe is designed for every interval of 500bp-2K, preferably one probe is designed for every interval of 1K. Further, probes were designed for all 79 exons. (Third-generation sequencers have long sequencing read lengths. For example, Pacbio’s sequel or Oxford Nanopore’s PromethION have a sequencing length of tens of K, and the genomic DNA in the sample to be tested is broken into fragments of about 5K in size, designed at intervals of 1K One probe, on average, each DNA fragment can bind 5 probes, which is enough to effectively capture the target fragment). The number of bases of the designed probe is between 50 and 130bp, the GC content is moderate, and there is no hairpin structure. It is further compared with NCBI and other databases for Blast comparison, and it is ensured that the probe avoids repetitive regions on the genome. These parameters are ...

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Abstract

The invention relates to the technical field of biology, in particular to a Dystrophin gene detection probe, a kit and a method. The invention also relates to a design method of the probe. The primerand the kit provided by the invention can be combined with a three-generation sequencing method to effectively sequence the human Dystrophin gene and analyze mutation in the human Dystrophin gene.

Description

[0001] This application claims the priority of the Chinese patent application submitted to the China Patent Office on March 29, 2019, the application number is CN 2019102488652, and the invention title is "Dystrophin gene detection probe, kit and method", the entire content of which is incorporated by reference in this application. technical field [0002] The invention relates to the field of biotechnology, in particular to a Dystrophin gene detection probe, kit and method. Background technique [0003] Dystrophinopathy (Dystrophinopathy) is a common neuromuscular disease, which is caused by a mutation of the Dystrophin gene on the X chromosome, resulting in a defect in the encoded dystrophin protein. According to age of onset, muscle pseudohypertrophy, course of disease and prognosis, it is divided into Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD). The former has severe symptoms and is a type of severe progressive muscular dystrophy. It starts at ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/6869C12N15/11
CPCC12Q1/6883C12Q1/6869
Inventor 高玉梅潘世让郎娜梁帆王洋汪德鹏
Owner BEIJING GRANDOMICS BIOTECH
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