Lily mottle virus double-antibody sandwich enzyme-linked immunosorbent assay kit and preparation method
A lily mottle virus, detection kit technology, applied in antiviral immunoglobulin, botanical equipment and methods, biochemical equipment and methods, etc., can solve the problem that there is no specific detection kit for LMoV, which is expensive and difficult Meet the needs of detoxification detection of flowers and edible lilies, and achieve good market prospects, high sensitivity and good specificity
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[0059] Example 1
[0060] 1. Preparation method of LMoV recombinant fusion protein PI200:
[0061] 1) Trizol method to extract total RNA from diseased lily leaves:
[0062] A. Weigh 40 mg of diseased lily leaves, put them in a mortar, and quickly grind them into powder in liquid nitrogen. After the liquid nitrogen volatilized, 1 mL of Trizol (Invitrogen) was added, and transferred to a 1.5 mL centrifuge tube, and allowed to stand at room temperature for 5 minutes.
[0063] B. Add 200μL of chloroform, mix upside down and mix several times, and leave it at room temperature for 2 to 3 minutes. Centrifuge at 12,000g for 10 min at 4°C.
[0064] C. Take the upper aqueous phase, add 500μL of isopropanol, mix well and let stand at -20°C for 20min, and centrifuge at 12,000g for 10min at 4°C.
[0065] D. Discard the supernatant, wash the precipitate with 1 mL of 75% ethanol, and centrifuge at 7,500 g for 5 min at 4°C.
[0066] E. Air-dry the precipitate and dissolve it with 20-40μL RNase-Free wat...
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