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
The seeding concentration may need adjustment to get a high-quality culture. Do a screening experiment with a broad 1:10 dilution series to find a suitable seeding concentration. See Optimize starter culture for more details.
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.
