Table 1.
Master Mix recipe
| Component | Stock Concentration | Volume Stock | Master Mix Final Concentrationa |
|---|---|---|---|
| Pyruvate Kinase | 1000 U/ml | 18.8 μl | 12.5 U/ml |
| Myokinase | 10000 U/ml | 1.9 μl | 12.5 U/ml |
| Lactate Dehydrogenase | 1000 U/ml | 18.8 μl | 12.5 U/ml |
| Phosphoenolpyruvate | 300 mM | 18.8 μl | 3.75 mM |
| NADH | 10 mM | 150 μl | 1000 μM |
| MOPS | 1 M | 375 μl | 250 mM |
| MgCl2 | 1 M | 28.1 μl | 18.74 mM |
| Citrate (0.1 × KM) | 25 mM | 4.1 μl | 67.5 μM |
| ahLys (0.1 × KM) | 25 mM | 5.9 μl | 98.75 μM |
| Water | 878.6 μl | ||
| Total Volume | 1500 μl |
a
Final concentration in master mix is 1.25 × concentration in the final reaction.