Open the tracking form and verify you also have a copy of the batch tracking sheet printed out:

• Step A tracking form
• Batch tracking sheet

Ensure agar plates are pre-poured

Pre-label the plates using only the numbers provided on the spreadsheet under heading ‘Sample_Label’

Get required glycerol stocks and keep on dry ice

Ensure to cross reference with spreadsheet before every inoculation

Create streak plate inoculum using inoculation loop from glycerol stock

Replace inoculation loop (after step I in the below figure) and dilute across the plate

Incubate at 37°C overnight

Pre-label tubes using numbers from spreadsheets

Transfer 5 ml of LB broth into a universal tube

Inoculate the LB broth with a single colony from the agar plate

Grow at 37°C overnight at 180 rpm in a shaking incubator

Place agar plates into fridge for storage