Reaction tube used in loop-mediated isothermal amplification technique and use method thereof
A loop-mediated isothermal amplification reaction technology, applied in the field of temperature amplification, can solve problems such as aerosol pollution in LAMP reaction, and achieve the effects of low cost, reduced pollution, and guaranteed detection.
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Embodiment 1
[0038] Such as image 3 As shown, the reaction tube of this embodiment is composed of a tube body and a tube cover. The lower part of the inner cavity of the tube is provided with a longitudinally extending partition separating it into two cavities A and B, wherein: cavity A There are 22.0 microliters of LAMP reaction solution and 0.5 microliters of Bst DNA polymerase inside, and the upper layer of the liquid is sealed with paraffin; the cavity B is filled with 2.0 microliters of LAMP reaction chromogenic solution, and the upper layer of the liquid is also sealed with paraffin, and the reaction Store the tube at -20°C.
Embodiment 2
[0039] The detection of embodiment 2 Mycobacterium tuberculosis
[0040] This embodiment adopts the reaction tube prepared in Example 1 (primers in the reaction solution are selected from a known primer group for detecting Mycobacterium tuberculosis), and detects Mycobacterium tuberculosis by LAMP, and a positive control group and a negative control group are provided at the same time. Process such as Figure 4 As shown, it specifically includes the following steps:
[0041] Take three reaction tubes prepared in Example 1, and add 2.5 μL each of negative control sample, positive control sample and sample to be tested respectively. When adding the sample, the tip of the pipette penetrates the protective liquid layer, and the sample is added to the large cavity of the Huafeng tube. Tightly cap and label the tube and move to the reaction area.
[0042] The above reaction tubes were reacted at a constant temperature of 65° C. for 1 h.
[0043] At the end of the reaction, shake ...
Embodiment 3
[0046] The detection of embodiment 3 Salmonella
[0047]This embodiment adopts the reaction tube prepared in Example 1 (primers in the reaction solution are selected from a known primer group for Salmonella detection) to detect Salmonella by LAMP, and a positive control group and a negative control group are provided at the same time. The process is as follows: Figure 4 As shown, it specifically includes the following steps:
[0048] Take three reaction tubes prepared in Example 1, and add 2.5 μL each of negative control sample, positive control sample and sample to be tested respectively. When adding the sample, the tip of the pipette penetrates the protective liquid layer, and the sample is added to the large cavity of the Huafeng tube. Tightly cap and label the tube and move to the reaction area.
[0049] The above reaction tubes were reacted at a constant temperature of 65° C. for 1 h.
[0050] At the end of the reaction, shake the reaction tube upside down once, and th...
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