TITLE: Making normal NGM
AUTHORS: Bonnie Evans

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">C. elegans is maintained in the laboratory on Nematode Growth Medium (NGM) agar which has been aseptically poured into petri plates using a peristaltic pump. NGM (rather than no peptone NGM) may also be used for imaging plates when screening bacterial strains. </div></div>

[STEPS]
?. [Pouring multi-well plates]
For pouring multi-well imaging plates:
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?. [Pouring Petri dishes]
For pouring maintenance/imaging Petri dishes:
{"blocks":[{"key":"c1ikd","text":"C. elegans is maintained in the laboratory on Nematode Growth Medium (NGM) agar which has been aseptically poured into petri plates. Smaller plates (35 mm diameter) are useful for mating or when using expensive drugs. Medium size plates (60 mm diameter) are useful for general strain maintenance, and larger plates (100 mm diameter) are useful for growing larger quantities of worms, such as for certain mutant screens. The NGM agar medium can be poured into petri plates easily and aseptically using a peristaltic pump. This pump can be adjusted so that a constant amount of NGM agar is dispensed into each petri plate. A constant amount of agar in the plates reduces the need for refocusing the microscope when you switch from one plate to another. ","type":"unstyled","depth":0,"inlineStyleRanges":[],"entityRanges":[],"data":[]}],"entityMap":[]}
?. [Pre-autoclave]
Book the autoclave via the notebook on top of the machine. It will take around 2 hours for a 500ml bottle and 2.5 hours for 1L or larger bottles.
?. [Pre-autoclave]
Take clean bottles and measuring cylinders from the glass kitchen - you will also need one bottle for the autoclave probe, which should be the size of the largest bottle you are autoclaving. Get sterile water from the media room.
?. [Pre-autoclave]
Measure out all the pre-autoclave reagents and add to the bottle. Use a new weighing boat and spatula for each reagent. Add water last and mix thoroughly.
?. [Pre-autoclave]
Add equivalent amount of water to the empty probe bottle.
?. [Pre-autoclave]
Make sure the lid of your media bottle is not screwed completely when placing it inside the autoclave machine. Label with autoclave tape (‘NGM agar Rm 5020’), and place over the lid to prevent it coming off in the autoclave.
?. [Using the autoclave]
Turn ON the autoclave and touch the screen
?. [Using the autoclave]
If 'Fill with water' is on the screen, fill up the autoclave with sterile water until it reaches the grill/'Water Level OK' appears on the screen.
?. [Using the autoclave]
Place the probe bottle inside the autoclave with the temperature probe inside.
?. [Using the autoclave]
Place the bottles in the autoclave and make sure that the cap is not screwed completely.
?. [Using the autoclave]
Check the waste flask is not too full
?. [Using the autoclave]
Close the door
?. [Using the autoclave]
(On the screen) LOCK --> START --> MEDIA 121X15 BALLAST --> Start
?. [Post-autoclave]
When autoclave is finished, UNLOCK the door
?. [Post-autoclave]
Remove the probe and media bottles using gloves (the bottles will be hot)
?. [Post-autoclave]
Leave the agar on a lab bench to cool to around 55°C, ie the bottle is cool enough to hold for a second with a gloved hand
?. [Post-autoclave]
Add the post autoclave reagents
?. [Post-autoclave]
Try not to shake the bottle too much while mixing to avoid air bubbles.
?. [Post-autoclave]
The agar needs to be warm to be poured without blocking the tubings, so try to pour as quickly as possible and if not poured immediately put the bottle on a waterbath set to 60C until being used.