Table 2.

CFE reaction mix compositions tested for the mycoplasma CFE platform, the standard concentration ranges for E. coli CFE are shown in the right column

Component	Mycoplasma CFE	Optimal E. coli CFE*
Mg-glutamate	0–25 mM	2.5–10 mM*
K-glutamate	60–120 mM	60–80 mM
DNA template	6–10 nM	5–8 nM*
Lysate buffer	1× S30B, 0.5× S30B, 75 mM HEPES pH 8.0	1× S30B, 0.5× S30B, deionized water
Lysate fraction	33–50 vol%	33 vol%*
Dilution solvent	1× S30B, 0.5× S30B, deionized water	Deionized water
RNase inhibitor	Recombinant RNase A/B/C inhibitors, RNAsecure®, PVSA	–

^*The optimal concentrations of Mg-glutamate, DNA template and lysate fraction varied from batch to batch. Therefore, we display concentration ranges in which we observed the best protein yields.