Research Article
Optimization of an In Vitro Transcription/Translation System Based on Sulfolobus solfataricus Cell Lysate
Table 1
Experimental conditions adopted for reactions with S30 S. solfataricus.
| | In vitro transcription adopted from [16] | In vitro transcription under our conditions | Coupled in vitro transcription and translation | In vitro translation |
| | KCl (mM) | — | 10 | 10 | 10 | | Tris-HCl (mM) | 50 (pH 8.0) | 20 (pH 6.8) | 20 | 20 | | Mg(OAc)2 (mM) | 25 | 20 | 20 | 20 | | ATP (mM) | 2 | 2 | 1.5 | 1.8 | | CTP (mM) | 1 | 1 | 1.5 | — | | GTP (mM) | 1 | 1 | 1.5 | 0.9 | | UTP (mM) | 0.6 | 0.5 | 1.5 | — | | (α-32P) UTP (μM) | 100 | 100 | — | — | | EDTA (mM) | 1 | — | — | — | | DTT (mM) | 1 | — | — | — | | Total tRNA (μg) | — | — | 3,3 | 3,3 | | S30 (μg) | 100–150 | 100–150 | 100–150 | 100–150 | | T (°C) | 60 | 70 | 70 | 70 |
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