Method and kit for detecting pathogenic bacterium of food source by test strip based on NASBA (Nucleic Acid Sequence Based Amplification)
A technology for food-borne pathogenic bacteria and test strips, which can be used in microorganism-based methods, microorganism determination/inspection, biochemical equipment and methods, etc.
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[0075] (1) Extraction of total RNA from Listeria monocytogenes (strain CMCC54007, purchased from Guangzhou Institute of Microbiology):
[0076] ①Grinding with liquid nitrogen, take about 5mL of bacterial liquid in a mortar, add a small amount of liquid nitrogen, and grind quickly. During the grinding process, add liquid nitrogen continuously until the grinding is complete. After grinding, transfer the cell lysate into a 1.5mL EP tube, then add 0.8mL Trizol to the grinder, blow the residual lysate on the mortar, and pour it into the EP tube.
[0077] ②Cover the lid, invert and mix 10 times, and let stand at room temperature for 5 minutes.
[0078] ③Add 0.2mL chloroform to the EP tube, close the cap, shake vigorously for 15 seconds to mix the liquid thoroughly, let stand at room temperature for 5 minutes, and then centrifuge at 12,000 rpm for 15 minutes.
[0079] ④ Transfer the supernatant to a new 1.5mL EP tube (approximately 0.5mL), add 0.5mL isopropanol, shake vigorously for...
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