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.
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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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