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Preloaded PCR reaction reagent for nucleic acid amplification and preparation method thereof

A technology for reaction reagents and nucleic acids, applied in the field of pre-installed PCR reaction reagents and their preparation, can solve the problems of increased transportation time and increased economic costs, and achieve the effects of improving stability, prolonging shelf life and ensuring stability.

Active Publication Date: 2021-04-06
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for the application of kits in remote areas, low-temperature transportation increases the economic cost and the transportation time will greatly increase

Method used

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  • Preloaded PCR reaction reagent for nucleic acid amplification and preparation method thereof
  • Preloaded PCR reaction reagent for nucleic acid amplification and preparation method thereof
  • Preloaded PCR reaction reagent for nucleic acid amplification and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Coronavirus (CoV) is a positive-strand RNA virus with a diameter of 80-120 nm. It is an important pathogen of humans and many animals and can cause a variety of acute and chronic diseases. There are many kinds of CoV, among which the genus α (including CoV-229E, CoV-NL63) and genus β (including CoV-OC43, CoV-HKU1, SARS-CoV, MERSCoV and SARS-CoV-2) that infect mammals are. SARS-CoV-2 is the seventh human-infecting coronavirus discovered so far, with a genome size of 29,891 nucleotides, including untranslated regions (UTRs) at both ends and a complete open reading frame gene ( openreading frame, ORF), which can encode 960 amino acids.

[0030] At this stage, the fluorescent PCR detection product of SARS-CoV-2 still adopts the conventional form of 1 tube of liquid reaction buffer + 1 tube of liquid enzyme. higher cost. The fluorescent PCR reagent freeze-drying method included in the present invention can well solve the above problems.

[0031] In order to verify the eff...

Embodiment 2

[0069] The PCR freeze-dried powder in the above-mentioned Example 1 was used as the control group, and the unlyophilized PCR reagent was used as the experimental group. The two groups were stored at room temperature for 48 hours and then tested by QPCR. The detection method was as in Example 1.

[0070] The results show

[0071]

Embodiment 3

[0073] Place the freeze-dried PCR preparation at 70°C, 0°C and -20°C to simulate the stability at high temperature, low temperature and extremely low temperature, respectively, and place it at each temperature for 3 hours to perform QPCR detection. The detection method is as in Example 1 .

[0074] The results show

[0075]

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Abstract

The invention discloses a preloaded PCR reagent for nucleic acid amplification and a preparation method of the preloaded PCR reagent. The preloaded PCR reagent comprises a PCR reagent, and is characterized in that the PCR reagent contains antifreeze protein and an antifreeze protein antibody. The reagent can be stored or transported at normal temperature.

Description

technical field [0001] The invention relates to a prepacked PCR reaction reagent for nucleic acid amplification and a preparation method thereof. Background technique [0002] PCR reaction (polymerase chain reaction) has played an important role in biological experiments since its invention, especially fluorescent PCR (qPCR) technology has played an increasingly important role in the field of microbial detection. Real-time fluorescent quantitative PCR is a PCR technology that can quantitatively monitor the copy number of the target gene. It is to add fluorescent dyes or fluorescent markers to the PCR system to combine with the amplification products to emit fluorescence, and to directly detect the fluorescent signal of the reaction system through the fluorescent signal detector. The relationship between the fluorescence intensity and the number of PCR cycles is plotted to quantitatively estimate the amount of PCR amplification and the amount of nucleic acid of the target gen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/686C12Q1/70C12R1/93
CPCC12Q1/686C12Q1/701C12Q2527/125C12Q2563/107C12Q1/6806
Inventor 曾子晏王薛婷
Owner XIAMEN UNIV
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