Preparing Agar Preparing Agar PlatesPlatesAs demonstrated by Andrew As demonstrated by Andrew CorcoranCorcoran
Use an appropriate flaskUse an appropriate flask
Use a flask that is Use a flask that is twice the volume of twice the volume of the media you will the media you will makemake
Typically a 1 L flask Typically a 1 L flask to prepare 0.5 L to prepare 0.5 L batchesbatches
Heat until there is a hint Heat until there is a hint of boilingof boiling
Avoid boiling over!Avoid boiling over! Agar will partially dissolve and get a little clearerAgar will partially dissolve and get a little clearer Wear a “Clavie”Wear a “Clavie”
Use autoclave tapeUse autoclave tape
Turns color after Turns color after heatingheating
Tear off tape and Tear off tape and apply to counter apply to counter (easier to write on it)(easier to write on it)
Include initials and Include initials and media typemedia type
Always place media into Always place media into autoclave trayautoclave tray
Secondary Secondary containmentcontainment
Spills in the Spills in the autoclave clog ventsautoclave clog vents
Operating autoclaveOperating autoclave
First rule – always make sure there is no First rule – always make sure there is no pressure in the autoclave!pressure in the autoclave!
Chamber pressure –
Should read zero
Jacket Pressure -
There should always be pressure here
Make sure previous cycle Make sure previous cycle is finishedis finished
Chamber pressure –
Should read zero
Autoclave status
Open the doorOpen the door
Lefty looseyLefty loosey Pins will retractPins will retract Crack 1 inch. Allow Crack 1 inch. Allow
excess steam to ventexcess steam to vent Open slowlyOpen slowly Remove contents Remove contents
carefully (contents carefully (contents may be superheated)may be superheated)
Pins
Place tray in center of Place tray in center of rackrack
Slide out metal shelf (avoid reaching into autoclave)Slide out metal shelf (avoid reaching into autoclave)
Shelves
Close the doorClose the door
Righty tightyRighty tighty Make sure pins Make sure pins
engageengage Door closed light will Door closed light will
be litbe lit Turn until hand tightTurn until hand tight
Press flask icon Press flask icon
Fast exhaust – Fast exhaust – ScissorsScissors
Fast exhaust with Fast exhaust with drying – Gowndrying – Gown
Slow exhaust - FlaskSlow exhaust - Flask
The control panelThe control panelFast exhaust
Fast exhaust/drySlow exhaust
Service – don’t touch
Start button
Cycles are pre-programmed. There is no need to change settings!
After autoclaving place After autoclaving place flask in 55 flask in 55 ooC waterbathC waterbath
Tape has darkened
Pouring the platesPouring the plates
Start by disinfecting the work areaStart by disinfecting the work area
Label plates on the Label plates on the bottombottom
Media typeMedia type DateDate Label the bag Label the bag
they will go inthey will go in
Remove lid and begin Remove lid and begin pouring plates from the pouring plates from the bottom of the stackbottom of the stack
After the first plate is filled After the first plate is filled cover and begin filling the cover and begin filling the second (and so on)second (and so on)
Avoid moving or jostling Avoid moving or jostling platesplates
Agar on the lid - faux pas!
Condensation will form on lid of plate
Stack the platesStack the plates
This prevents condensation on most platesThis prevents condensation on most plates Also provides more work roomAlso provides more work room