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Primer combination and reagent kit for complete mitochondrial genome detection

A whole genome, primer combination technology, applied in the field of biotechnology detection, can solve problems such as amplification failure

Active Publication Date: 2017-05-31
北京圣谷智汇医学检验所有限公司
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Problems solved by technology

[0011] 2. The mitochondrial genome sequence is highly polymorphic. If primers are designed at polymorphic positions, the amplification will fail due to the mismatch between the primer and the template sequence.

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  • Primer combination and reagent kit for complete mitochondrial genome detection
  • Primer combination and reagent kit for complete mitochondrial genome detection

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Embodiment Construction

[0092] The term "primer" as used herein can refer to a short linear oligonucleotide that hybridizes to a target nucleic acid sequence to prime the synthesis of a primer extension product under suitable conditions. In the polymerization reaction system, there can be one or more primers. The term "primer pair" as used herein refers to a set or pair of primers, including a 5' sense primer (also referred to as a "forward primer" or "upstream primer") that hybridizes to the complement of the 5' end of a target nucleic acid sequence. ", which can be represented by the abbreviation "F"), and a 3' antisense primer (also known as "reverse" or "downstream", which can be represented by the abbreviation "R") that hybridizes to the 3' end of the target sequence.

[0093] As used herein, the term "amplification" refers to an in vitro method of increasing the copy number of a target nucleic acid sequence in a sample. The amplification reaction typically consists of multiple rounds of repeate...

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Abstract

The invention relates to a primer combination and a reagent kit for complete mitochondrial genome detection. A PCR and Sanger sequencing technology is used for detecting a complete mitochondrial genome for auxiliary diagnosis of a mitochondrial disease caused by mtDNA (mitochondrial deoxyribonucleic acid) mutation. Six specific primer pairs are used for amplification of the whole mitochondrial genome first; each amplification product is 2700-3500bp in size; 200bp or more parts of adjacent amplification segments are overlapped; then, 21 sequencine primers are used for direct sequencing of the amplification products; the sequencing range covers the whole mitochondrial genome. With the adoption of the primer combination, the whole mitochondrial genome can be detected by 6 amplification reactions and 21 sequencing reactions; the detection workload is greatly reduced; moreover, the detection cost is also greatly lowered.

Description

technical field [0001] The invention relates to the field of biotechnology detection, in particular to primers and a kit for mitochondrial whole genome detection. Background technique [0002] Mitochondrial disease (mitochondriopathy) is a group of multi-system diseases or tissue-specific diseases caused by defects in mitochondrial metabolic enzymes caused by genetic defects, resulting in ATP synthesis disorders and insufficient energy sources. Mitochondria are the energy-producing organelles within cells. It is present in every cell in the body except red blood cells. The main function of mitochondria is to provide the energy needed by cells - adenosine triphosphate (ATP). Mitochondria have their own genetic control system, which is also controlled by nuclear DNA. Once the mitochondrial DNA (mtDNA) is mutated, it will cause the enzymes or carriers necessary for encoding the mitochondrial oxidative metabolism process to be impaired. Glycogen and fatty acids cannot enter t...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6869C12Q1/6883C12Q2600/156C12Q2531/113C12Q2535/101
Inventor 周仲春叶建伟张捷崔丹刘棒余应襄
Owner 北京圣谷智汇医学检验所有限公司
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