LAMP technology-based primer, kit and detection method for detecting Plum pox virus
A plum pox virus, technical detection technology, applied in the biological field, can solve the problems of easy degradation, false positive, cumbersome RNA extraction, etc.
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Embodiment 1
[0112] Kit of the present invention:
[0113] 1. Product specification: 50 times
[0114] 2. Features:
[0115] Loop-mediated isothermal amplification method loop-mediated isothermal amplification method, LAMP is a new gene amplification method, which uses only one enzyme to amplify at a constant temperature, and uses 6 primers that can recognize 6 specific sequences of the target DNA, so the specificity High, and the amplification efficiency is very high, it can be amplified in a short time, and the reaction result can be judged visually.
[0116] This kit adopts the loop-mediated isothermal amplification method, and uses ribonucleic acid RNA as a template to directly carry out a simple kit for reverse transcription loop-mediated isothermal amplification. The sample to be tested and the reagents in this kit need to be mixed according to the requirements, and then placed at a certain temperature at 63°C for about 1 hour to react, and the result can be judged from the amplifi...
Embodiment 2
[0140] The present invention detects the detection method of plum pox virus, comprises the following steps:
[0141] A. Using nano magnetic beads to extract the RNA of the sample to be tested;
[0142] The immune nanomagnetic beads are produced by the following process:
[0143] ① Preparation of nanomagnetic beads
[0144] Take 5.2gFeCl respectively 3 ·6H 2 O, 2.7g FeSO 4 ·7H 2 O. Dissolve 0.85mL of 12.1moL / L concentrated hydrochloric acid in 200mL of water, ultrasonically deoxygenate, and then drop the above solution into 250mL, 0.75moL / L NaOH solution; stir at 80°C, after the reaction is over, use a magnetic separator to separate the resulting precipitate Separated from the reaction medium, washed 3 times with deionized water and 2 times with absolute ethanol, and made into 5g / mL Fe 3 o 4 Magnetic nanoparticles absolute ethanol solution; take 25mLFe 3 o 4 Magnetic nanoparticle absolute ethanol solution, then diluted with absolute ethanol to 150mL, ultrasonically osc...
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