TITLE: Biomek 96 well plating
AUTHORS: William J Buchser, jwaligor

[DESCRIPTION]
Randomized plating of cells using Biomek. 

[STEPS]
SECTION: Creating Plate Layouts in FIVTools
1. Open FIVTools and click "Layouts."

SECTION: Creating Plate Layouts in FIVTools
2. Edit "Exp Name" to the correct FIV Number (make sure to keep "FIV" in the name)
1. Click once on the “Export Folder” bar so it updates the location
SECTION: Creating Plate Layouts in FIVTools
3. Choose the desired number of 96-well plates by typing 1, 2, 3, or 4 in "Count."
Select “96 Mote” to exclude plating from the outermost wells
Select “384 Full” for a 384 well plate
If the layout is not standard, adjust the “First Row”, “First Column”, “Last Row”, and “Last Column” boxes appropriately
If doing an antibody plate, check the “Ab Plate” box and:
        - Make “Count” = 0 for one antibody plate
        - Make “Count” = 2 for two 96-well plates and an antibody plate

SECTION: Creating Plate Layouts in FIVTools
4. Change “Cells per Well” to desired plating density.
SECTION: Creating Plate Layouts in FIVTools
5. Copy experimental condition names (often genotypes, dosages, G1/G2, etc.) and click “PASTE Names” to auto-fill the 12-well source plate.
“Name” can also be manually filled
The “Well” column can only have row/column combinations of a 12-well plate (A-C, 1-4)
“Relative Representation” allows you to assign different weights for representation in the 96-well plates
Use the underscore symbol to have a cell type only show up in the antibody plate layout (for example: “_cell type”)
SECTION: Creating Plate Layouts in FIVTools
6. Click "Fill Plates"
1. Try to make sure the empty wells are not on any edges and if doing multiple plates that they cannot be flipped or rotated to the same location as another plate. 

SECTION: Creating Plate Layouts in FIVTools
7. Go into the “4 Mapping” folder of the respective experiment and make sure there is a PlateMap, PlatingSetup, and script. 
SECTION: Setting up Biomek
8. Find the correct method. 
 
SECTION: Setting up Biomek
8.1. The algorithm will look like this:
 
SECTION: Setting up Biomek
9. Set up the deck in the configuration shown
 
SECTION: Setting up Biomek
9.1. Make sure the two tip rows closest to the front are full in the “tip_box” and the “empty_1” and “empty_2” are completely empty.
SECTION: Setting up Biomek
9.2. Only use the number of 96-well plates needed for your experiment and place them in order starting with “0.”

    - Keep the lids on the plate until you are ready to begin the Biomek program.
SECTION: Setting up Biomek
10. Open the appropriate experiment folder, go to “4 Mapping” and open the “FIVXXX_script” file.
SECTION: Setting up Biomek
11. Copy the entire script.
SECTION: Setting up Biomek
12. Go to the “Scripted Let” step in the algorithm and paste your entire script. 

    - Scroll up to the top to make sure the correct experiment number and an appropriate time stamp are there
SECTION: Setting up Biomek
13. Go back to the “4 Mapping” folder and drag and drop the “FIVXXX_PlatingSetup” file onto and EXCEL sheet (DO NOT OPEN AS TEXT FILE). 
SECTION: Setting up Biomek
14. After creating cell suspensions for each source condition and obtaining counts, input their counts into the appropriate positions of the “Cells/uL” column: 
 
SECTION: Setting up Biomek
15. With a new 12-well plate, set up the Source Plate by adding the volumes in the “uL Media” and “uL Cells” columns into the appropriate wells: 
 
SECTION: Setting up Biomek
16. Place the Source Well in the correct position on the deck,open the lids to all plates, and close the front door. 
SECTION: Setting up Biomek
17. Press “Run” (the green arrow) and confirm the configuration in the pop-up box.