Crystal Violet for Plaque Assay

Marion Freistadt mfreis at
Sun Apr 24 11:54:18 EST 1994

>I am doing plaque assays for polio virus.  After I remove the agar
>from my 6-well TC plates, I would like to stain them with a
>solution of crystal violet.  Does any one have a recipe for CV
>for use in a plaque assay?  I have tried several microbiology
>manuals and each one seems to use a slightly different mixture.
>The tissue culture manuals that I have do not even mention CV.
>Thanks in advance for any and all help.
>Robert Citek

11. Crystal violet stain:  stock=1 g. crystal violet
                                                 99ml 20% EtOH
                       working soluion= 20 ml stock
                                                   40 ml 95% EtOH
                                                   150 ml distilled water
11. First apply 2 ml of 20% TCA to each well for 5 min. to fix.  Pour off
TCA.  Remove agar by ringing with spatula and flipping out into biohazard
12. Add 2 ml of crystal violet working solution to each well for 30 min. at
room temp.  Rinse gently with water.  All waste goes into virus bucket 
with bleach to be discarded.  Dry and count plaques.
Marion Freistadt, Ph.D.
Department of Microbiology, Immunology and Parasitology
Louisiana State University Medical Center
1901 Perdido St.
New Orleans, LA. 70112

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