Inoculate liquid culture

Use the subcultivation calculator tool to determine the volume needed to seed the growth medium. A typical seeding concentration for an overnight culture is 1 x 10⁶ cells/mL.

Use Optiseeding workflow to determine suitable seeding concentration

The seeding concentration may need adjustment to get a https://github.com/sbtinstruments/help--mpd/blob/main/encyclopedia/bacterial-growth/high-quality-culture.md. Do a screening experiment with a broad 1:10 dilution series to find a suitable seeding concentration. See https://github.com/sbtinstruments/help--mpd/blob/main/workflows/optimize-bioprocesses/identify-optimal-inoculum-size/README.md for more details.

1

Open the subcultivation calculator

Open the subcultivation calculator tool in a browser window.

2

Calculate inoculation volume

• Fill in cells/mL of the capture vial in the first field.

• Add volume information in the second field. In this case it is 40 mL

• Set the desired inoculation concentration in the final field. In this case 1 × 10⁶ cells/mL

• Click Calculate V_add to get the result.

3

Add starter culture to growth curve flask

Transfer starter culture to growth curve flask and swirl aggressively to disperse the inoculum.

Record the inoculation time. Record it on the autoclave tape as a backup, ensuring you have a reference if you forget to document it elsewhere.

Inoculation of shake flask at well-defined concentration. In this video the 50 mL vial with the red cap represents the capture vial.

Summary

The overnight culture is inoculated at a well-defined starting concentration of 1 ^_x 10⁶ cells/mL. Next step is to Determine concentration after incubation.