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a. Suspend the freshly cultured bacteria with distilled water or PBS, and adjust the
bacterial concentration to 1 x 108/ml. It must be noted that zoonosis pathogens must
be treated carefully, and inactivation is the best choice.
b. Add 100ul 5% glutaraldehyde solution (95ml 0.1mol/L NaHCO3 and 5ml 25%
glutaraldehyde solution) into each hole. Incubate for 2 hours at 37 , and wash for 3
times with distilled water. Add 50ul bacterial suspension from the last step into each
hole, and dry the late at 3756. Then, add 200ul blocking solution into each hole,
and maintain overnight at 4 or seal up for 2 hours at 37.
c. Step b can also be replaced by this way. Add 50ul bacterial suspension into each
hole. Dry at 37 -56 , then maintain at room temperature for 15 minutes with -20
pre-cooled anhydrous methanol and wash for 3 times with distilled water. Add 200ul
blocking solution, and maintain at 4 overnight or at 37 for 2 hours.
d. Wash 3 times with washing solution. Maintain the coated plate at -20 or 4 for
later use.
a. Culture the cells with a conventional manner. Inoculate virus, harvesting infected
cells and uninfected cells. Count obtained cells. Make suspension of appropriate
concentration with PBS.
c. Add 100ul cell suspension from step a or step b to each hole, making the whole cell
number of each hole 5104. Separate the supernatant at 1500r/min for 15 minutes.
Dry at room temperature or blow-dry, and fix for ten minutes with
acetonemethanol (1: 1) at 4. Store at 4 or -20 for later use.
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