54 Life Technologies
™
| Selection and Stable Pool Considerations
In Phase II selection, use a methotrexate stock with a concentration of
1.0 mM. Calculations for the amount of methotrexate and puromycin
have already been done for you—see Figure 10. 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 50P/1,000M condition
25 mL x 5 µL = 125 µL puromycin added to flask
25 mL x 1 µL = 25 µL 1.0 mM methotrexate added to flask
Selection and Stable Pool Considerations
Tip:
