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How-to

Inoculate liquid culture

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

Use Optiseeding workflow to determine suitable seeding concentration

1

Open the subcultivation calculator

Open the subcultivation calculator tool in a browser.

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

You have now inoculated the overnight culture at a well-defined starting concentration of 1 x 106 cells/mL. Next step is Determine concentration after incubation.

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