TITLE: Dispensing agar into multiwell plates
AUTHORS: Ida Barlow

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span>Protocol for pouring agar into 96 well plates using Integra VIAFILL dispenser. Agar should be prepared in advance, and kept in 60</span><span style = "vertical-align:super;">o</span><span>C waterbath until ready to dispense, and whilst dispensing. The X, Y, Z positions for dispensing can be adjusted according to the multiwell plates being used. </span></div></div>

[STEPS]
?. [Configure Integra VIAFILL]
Prepare a 250ml bottle of hot milliQ water in the microwave and keep in the waterbath along with the agar. The water is important to have on hand in case of tubing blockages.
?. [Configure Integra VIAFILL]
Insert large cassette into the machine
?. [Configure Integra VIAFILL]
Configure X, Y, and Z settings for the multiwell plate by clicking on tool symbol -> stage alignment.
For UNIPLATE96SQWLF 650U:X = 95.6Y = 4.2Z = -22.5For 48WP:
?. [Configure Integra VIAFILL]
Put the plate into the stage and then press 'Move' so that the plate moves so that it is under the dispensing cassette.
?. [Configure Integra VIAFILL]
Use the up and down arrows to move the pipette tip so that they hover just over the plate and make note of the height (this will be entered into the dispensing program at a later step). Press 'Fast/Slow' button to switch between fast and slow movements.
?. [Configure Integra VIAFILL]
Use the X, Y arrows to move the plate so that the pipette tips are centered in the middle of column 5.
?. [Configure Integra VIAFILL]
Save all settings.
?. [Configure Integra VIAFILL]
Exit settings by pressing the back button
?. [Dispensing Agar]
Press on the program you wish to use (see later for configuring your own program)
?. [Dispensing Agar]
Make sure that the correct cassette is listed and change if necessary
?. [Dispensing Agar]
Select the volume you wish to dispense
For 96WP:200 μL
?. [Dispensing Agar]
Select 'set height' and set the appropriate height for tip height (usually all the same)
?. [Dispensing Agar]
Place the end of the tubing from the casettee into the agar that is being kept warm in the water bath
?. [Dispensing Agar]
Press 'Prime' to prime the tubing and allow to finish so that agar flows from the pipette tips.
IMPORTANT:Once the agar is in the tubing it is important to act quickly to avoid agar solidifying and causing blockages. If you are particularly concerned about agar cooling in the tubing, wrap the tubing in aluminium foil to keep hot.
?. [Dispensing Agar]
Place a clean plate in the stage
?. [Dispensing Agar]
Press run and then plate should fill with agar
?. [Dispensing Agar]
Repeat steps 11-12 until all the plates have been filled.
Little drops of agar can solidify on the tip ends. It is often good to remove these drops using a pipette tip every few runs so that blockages do not occur.
?. [Cleaning the cassette]
'Reverse prime' all the agar
?. [Cleaning the cassette]
Place the tubing ends into the hot water.
?. [Cleaning the cassette]
Prime so that the water runs through and clears all the agar
?. [Cleaning the cassette]
Reverse prime to remove the water
?. [Cleaning the cassette]
Release tension from the tubing and remove cassette
?. [Cleaning the cassette]
Double wrap the cassette in aluminium foil for autoclaving