Overview¶
Small Molecule Mix (SMix) contains the 31 small molecules of Base Cytosol: buffers and salts, the four rNTPs, the 20 canonical amino acids, stabilizers, and cofactors. With Protein Mix (PMix), ribosomes, and tRNA, it makes Base Cytosol.
Reference Composition¶
Composition of the Small Molecule Mix (SMix) at 3.33× concentration. Magnesium acetate is 25 mM here, which contributes 7.5 mM to the reaction.
| Category | Molecule | Concentration | Units |
|---|---|---|---|
| Buffers | HEPES-KOH (pH 7.6) | 166.67 | mM |
| Potassium Glutamate | 333.33 | mM | |
| Magnesium Acetate | 25 | mM | |
| Nucleotides | rATP | 6.67 | mM |
| rGTP | 6.67 | mM | |
| rCTP | 3.33 | mM | |
| rUTP | 3.33 | mM | |
| Amino Acids | Each (Ala–Val) | 1.00 | mM |
| Stabilizers | Spermidine | 6.67 | mM |
| TCEP-HCl | 3.33 | mM | |
| Cofactors | Creatine Phosphate | 66.67 | mM |
| Folinic Acid | 0.067 | mM |
Processes¶
Make Amino Acid Mix — the equimolar amino acid stock that SMix takes
Credits¶
Reformulated by Yemo Ku and Jon Calles (b.next) from the PURE system, developed by the Ueda Lab (Shimizu et al., 2001).
- Shimizu, Y., Inoue, A., Tomari, Y., Suzuki, T., Yokogawa, T., Nishikawa, K., & Ueda, T. (2001). Cell-free translation reconstituted with purified components. Nature Biotechnology, 19(8), 751–755. 10.1038/90802