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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 conditionsCoupled in vitro transcription and translationIn vitro translation

KCl (mM)101010
Tris-HCl (mM)50 (pH 8.0)20 (pH 6.8)2020
Mg(OAc)2 (mM)25202020
ATP (mM)221.51.8
CTP (mM)111.5
GTP (mM)111.50.9
UTP (mM)0.60.51.5
(α-32P) UTP (μM)100100
EDTA (mM)1
DTT (mM)1
Total tRNA (μg)3,33,3
S30 (μg)100–150100–150100–150100–150
T (°C)60707070