Pseudomonas fluorescens LAMP (loop-mediated isothermal amplification) detection agent and kit
A Pseudomonas fluorescens detection reagent technology, applied in the field of Pseudomonas fluorescens LAMP detection reagents and kits, to achieve high amplification efficiency, simple operation, and short detection time
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[0017] Below in conjunction with embodiment the present invention is described in further detail.
[0018]1. Design primers: select the cDNA of Pseudomonas fluorescens as the target, and use PRIMEREXPLORER V4 software to design primers (including the five F3, B3, Y-LB, FIP, and BIP). Bioengineering Limited completed. Compare the primers with the target, perform routine screening tests and analysis, and obtain specific, sensitive, and stable 5 primers for the above-mentioned F3, B3, Y-LB, FIP, and BIP; After optimizing the experiment, determine the reaction conditions with the best amplification efficiency and specificity.
[0019] 2. Reaction temperature optimization: Design a gradient LAMP reaction program, the reaction temperatures are: 61°C, 63°C, 65°C, and 63°C is optimized as the best reaction temperature among these temperatures.
[0020] 3. Suitable Mg 2+ Concentration selection: Mg in the reaction system 2+ (2.5 mM) was added in the following order: 0 μL, 2...
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