Hello,
I want to determine the infectious titer of an HSV strain on vero cells in a 96 wells plate. This is the first time i use the TCID50 test, so i have some basic questions:
-how many cells do you plate? You're expected confluence?
-the medium you use: 2%serum DMEM?
-Do you eliminate the old medium before adding your viral diltution? Or it won't make any difference bcz you conserve the same amout of total infectious particles?
-How many days in general do you wait for final reading?
-An infected well at 1 or 2% is considered +?
thank you,
Plate cells so that wells (of 96 well plate) are at 50% confluency in 2% serum DMEM. You do not have to remove medium, just add virus (I generally do a series of virus dilutions from 10E-3 to 10E-14). I generally wait 1 week to analyze plate, and yes any infected well (even at 1 or 2%) is considered positive. Good luck.
Hello
i agree with Susan D Kraner , this a good way
Good Luck
http://www.jove.com/visualize/abstract/24817544/attachment-penetration-acyclovir-resistant-herpes-simplex-virus-are
http://www.revistas.usp.br/bjps/article/viewFile/10663/12431
https://www.labome.com/method/Methods-for-Rapid-Virus-Identification-and-Quantification.html
Dear Michel
- It will be good if you use 1000 to 100,000 cells per each well
- 2% serum DMEM is good
- Dont need to discard old media
- 7 to 14 days
- Infected cells will be consider as positive
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