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Published byChastity Heath
Modified about 1 year ago
Preparing Agar Plates As demonstrated by Andrew Corcoran
Use an appropriate flask Use a flask that is twice the volume of the media you will make Typically a 1 L flask to prepare 0.5 L batches
Measure out the water
Add a spin bar and spin
Weigh out the appropriate amount of media
Add the powder Some powders are very fine
Make sure all the powder gets hydrated Wad of dry media
Heat until there is a hint of boiling Avoid boiling over! Agar will partially dissolve and get a little clearer Wear a “Clavie”
Remove the spin bar with the wand Wand Spin Bar
Apply an aluminum foil lid
Use autoclave tape Turns color after heating Tear off tape and apply to counter (easier to write on it) Include initials and media type
Apply to flask
Always place media into autoclave tray Secondary containment Spills in the autoclave clog vents
Operating autoclave First rule – always make sure there is no pressure in the autoclave! Chamber pressure – Should read zero Jacket Pressure - There should always be pressure here
Make sure previous cycle is finished Chamber pressure – Should read zero Autoclave status
Open the door Lefty loosey Pins will retract Crack 1 inch. Allow excess steam to vent Open slowly Remove contents carefully (contents may be superheated) Pins
Place tray in center of rack Slide out metal shelf (avoid reaching into autoclave) Shelves
Close the door Righty tighty Make sure pins engage Door closed light will be lit Turn until hand tight
Press flask icon Fast exhaust – Scissors Fast exhaust with drying – Gown Slow exhaust - Flask
The control panel Fast exhaust Fast exhaust/dry Slow exhaust Service – don’t touch Start button Cycles are pre- programmed. There is no need to change settings!
Steam pressure will rise
After autoclaving place flask in 55 o C waterbath Tape has darkened
Pouring the plates Start by disinfecting the work area
Stack plates 4 high (right side up)
Label plates on the bottom Media type Date Label the bag they will go in
Wipe away water from flask
Remove lid and begin pouring plates from the bottom of the stack
After the first plate is filled cover and begin filling the second (and so on)
Avoid moving or jostling plates Agar on the lid - faux pas! Condensation will form on lid of plate
When done, put hot water in flask
Pour into special trash can
Stack the plates This prevents condensation on most plates Also provides more work room
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