52 Life Technologies
™
| Selection and Stable Pool Considerations
In Phase I selection, use a methotrexate stock with a concentration of
0.1 mM. The lower concentration will help with more accurate pipetting
of the selection agent. Calculations for the amount of methotrexate and
puromycin have already been done for you—see Figure 9. The calculated
numbers represent how much selection agent you should add per
milliliter of fresh medium.
For example: 25 mL of fresh medium for the 20P/200M condition
25 mL x 2 µL = 50 µL puromycin added to flask
25 mL x 2 µL = 50 µL 0.1 mM methotrexate added to flask
Selection and Stable Pool Considerations
Tip:
