Overview¶
The deGFP Reporter Module produced deGFP, a green fluorescent protein.
Schematic
Designs

A schematic representation of PURE converting template DNA into a fluorescent reporter.
| Name | Length (bp) | File |
|---|---|---|
pOpen-pT7-deGFP | 2500 | path |
Cytosols¶
Reference Composition¶
This composition was evaluated in the DevNote by Yadav (2025)
| Component | Stock Concentration | Final Concentration in Reaction |
|---|---|---|
| PMix | 15 mg/mL | 1.8 mg/mL |
| SMix | 4X | 1X |
| Ribosomes | 10 M | 1.8 M |
| tRNA | varies | 3.5 mg/mL |
| pOpen-deGFP | varies | 3 nM |
| Magnesium acetate | 200 mM | varies |
Expected Behavior¶
Kinetics
Endpoint

Translation kinetics of Cytosol and PURExpress reactions using two different pOpen-deGFP DNA preps. Cytosol w/o DNA refers to the Cytosol reaction lacking the pOpen-deGFP template. Data from Yadav (2025).

Final protein yields of the reactions measured at steady state. Data from Yadav (2025).
Protocols¶
Cells¶
Reference Composition¶
Cytosol
Membrane
Outer Solution
| Component | Input concentration | Unit | Final concentration | Unit | Volume for one reaction [µL] |
|---|---|---|---|---|---|
| SMix | 3.33 | × | 1 | × | 12 |
| PMix | 15 | mg/mL | 1.80 | mg/mL | 4.8 |
| Ribosomes | 10 | µM | 1.8 | µM | 7.2 |
pOpen-deGFP DNA | 124 | nM | 3 | nM | 0.95 |
| tRNA | 35 | mg/ml | 3.5 | mg/ml | 4 |
| Magnesium acetate | 200 | mM | 8 | mM | 1.6 |
| Optiprep | 1.32 | mg/uL | 0.043 | mg/uL | 1.33 |
| RNase Inhibitor | 40000 | U/mL | 2000 | U/mL | 2 |
| Water | 6.12 | ||||
| Total volume [µL] | 40 |
| Component | Target Percentage (%) | Molecular Weight (g/mol) | Stock concentration (mg/mL) | Volume to add (uL) |
|---|---|---|---|---|
| POPC | 70 | 760.076 | 25 | 162.17 |
| Cholesterol | 29.95 | 386.654 | 50 | 17.65 |
| Liss-Rhod PE | 0.05 | 1301.71 | 1 | 4.96 |
| Component | Concentration |
|---|---|
| Glucose | 850 mM |
Expected Behavior¶
Coming soon!
Protocols¶
- Yadav, S. (2025). First Nucleus Cytosol Testing: Evaluation of Nucleus Cytosol v0.5 Through deGFP Expression. 10.63765/fppr8928