Integrating PPK Module in PURE Cells

This DevNote implements the combined polyphosphate kinase (PPK) and creatine kinase-create phosphate (CP) system in the PURE Cell. To extend the results of from bulk reaction to a synthetic cell context, we compare the performance of the PPK, CP, and combined PPK + CP system in bulk reactions supplemented with either PEG4k or Optiprep, which is commonly used in the preparation of synthetic cells prepared by the emulsion transfer method. We find that optiprep resulted in overall reduced performance compared with PEG4k and unsupplemented reactions. Finally, we compared the performance of the PPK, CP, and PPK + CP systems in PURE Cells and find PPK + CP system again displaying the strongest signal, consistent with the trends observed in bulk measurements.