Agar Plate Making

Need 20ml LB/Agar for each plate

Read bottle for agar concentration

Ie 40g/L

If you want to make 20 plates you’ll need a total volume of 400ml (make a little more for error) 425ml

40g/L X .425L = 17g LB + 425ml diH2O (to erlenmeyer flask)

Autoclave (liquid protocol)

Remove from autoclave and cool to a temp where it is comfortable to touch flask (don’t want it to solidify)

Add antibiotic ( need 1X in total; start with 1000X stock)

If you have a total volume of 425ml use M1V1=M2V2 to figure out how much antibiotic to add

(1000X)(Yml)=(1X)(425ml)

Y=.425ml remember to convert to ul

.425ml X 1000ul/1ml= 425ul

Mix by swirling

Add approximately 20ml to each plate

Mark plate with appropriate color that corresponds to the particular antibiotic that was added

Colors:

Carbenicillin-Green

Kanamycin-Red

Chloramphenicol-Blue

Gentamycin-Purple

See antibiotic concentration page here