From agar plate colony

Get an active culture using a fresh colony from an agar plate.

Skill level	Time to complete	Hands-on time	HW requirement
Intermediate	16-24 hours (E. coli)	3 hours	7.6a

A fresh colony from an agar plate is a good starting point to ensure a single-species (axenic) culture. The standard workflow uses overnight incubation of the liquid culture, but it is often possible to get a same-day starter culture from an agar plate (see hint box).

Illustration of workflow. A fresh colony is collected and transferrred to a capture tube. Cell clumps are disaggregated using bead-beater or vortex mixer. Finally, a sample of capture tube is diluted and subjected to BactoBox® measurement.

Get a same-day starter culture

• If you want a same-day starter culture, try using higher seeding concentration, e.g. 5× 10⁷ cells/mL.

• Check the liquid culture frequently for increase in cell concentration, e.g. for each 2 hours.

• We recommend that - prior to use - the active starter culture should have cell concentration is at least tenfold higher than at T₀.

The protocol assumes that you have already used the streak plate method to get fresh colonies on an agar plate. The steps are outlined below.

1. Get things ready

2. Collect and disperse bacteria from agar plate

3. Determine cells/mL in harvest tube

4. Inoculate liquid culture

5. Determine concentration of overnight culture

Next, the get things ready section will provide you with a check-list of necessary items.