A kit for detecting Listeria monocytogenes, Vibrio parahaemolyticus and Salmonella typhimurium and its preparation method
A technology of Salmonella typhimurium and Vibrio hemolyticus, which is applied in the field of kits, can solve the problem of long-lasting antibodies and achieve the effects of shortened detection time, small coefficient of variation, and good stability
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[0045] The present invention also provides a preparation method of a kit for detecting Listeria monocytogenes, Vibrio parahaemolyticus and Salmonella typhimurium, comprising:
[0046] Step 1: Preparation of self-priming valve-separated chips
[0047] 1) Preparation of the mold
[0048] Use CorelDRAW X8 to design the channel and chamber pattern of the self-priming valve compartment chip, and then print it on a transparent film as a mask. Considering the height of the chamber, it is preferable to coat the silicon wafer with two layers of negative light For photoresist, use Spincoat G3P-8 spin coater to apply negative photoresist on clean and dry silicon wafers. The operating conditions of the spin coater are preferably 2000-3000rpm, and the time is 30s-1min, and then repeat this process again. step to form the second layer, then place the silicon wafer on a "h49-25c type 4" single-sided lithography machine under the mask, before exposure, adjust the position of the mask so that...
Embodiment 1
[0058] Example 1 Preparation of self-priming valve partitioned chip
[0059] The mold was treated with trimethylchlorosilane (MACKLIN, China) for 10 min to prevent polydimethylsiloxane from sticking. The polydimethylsiloxane component Prefix A and Crosslinker B (#RTV 615, Momentive, USA) were then mixed in a mass ratio of 10:1, air bubbles removed and poured into a mold. After the chip was at rest for about 1 min, the air bubbles generated in the PDMS during pouring were removed. Thread the bolt and nut flush on one end, then place the flush end down at the valve position of the mold, taking care to avoid introducing air bubbles into the polydimethylsiloxane that could affect chip performance. Then placed on a heating plate and baked at 90° C. for 30 min. After the polydimethylsiloxane layer was completely cured, it was carefully peeled off from the mold, and a 1.2 mm hole punch was used to punch the sample hole. Finally, plasma pretreatment was performed on the channel surf...
Embodiment 2
[0060] The preparation of embodiment 2 graphene oxide
[0061] Add 1.0g graphite powder to 100mL H 2 SO 4 and 46 g of sodium nitrate, and stirred in an ice bath for 30 min. With vigorous stirring, 3.0 g of potassium permanganate was slowly added to the above mixture (the temperature was kept at 0°C, but completed within 10 min) and stirred for 2 h. Continue to stir at 40°C for 2h, then heat to 150°C and stir for 3h. The reaction system was terminated by adding 200 mL of deionized water. H was then neutralized with NaOH in an ice bath 2 SO 4 , adjust the pH to 6. The resulting pale yellow solution was filtered through a 0.22 μm ion filtration membrane to remove large particles, and then dialyzed in a dialysis bag (MWCO 1000da) for 3 days to remove salt to obtain graphene oxide.
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