User guide

Clonal selection by limiting dilution, continued
Cell counting and
dilution
1. Label five 50-mL conical tubes “1” through “5”.
2. Pipette 5 to 10 mL of your transfected pool CHO DG44 cells into the 50-mL
tube labeled “1”.
3. Determine the viable cells/mL accurately using your method of choice.
4. Serially dilute the cells to a final concentration of 1000 cells/mL using growth
medium (CD OptiCHO
™
Medium supplemented with 8 mM L-glutamine) as
shown in figure below to yield a seeding density of 1 cell/well. If a different
cell/well seeding density is desired, adjust accordingly in the final dilution
step.
Note: Mix cells gently after each dilution by inverting the capped tube 5 to 6 times.
Avoid foaming.
Example of cells serially diluted in CD OptiCHO
™
Medium with an initial count
of 2.77 × 10
6
viable cells/mL
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