Fig. 2

Cell-free TxTl of GFP from dsDNA linear template with different promoters. a cell-free TxTl synthesis of eGFP, with two top candidate promoters, end point fluorescence measured after 8-hour reactions. b: RT-qPCR measurement of mRNA abundance in TxTl GFP translation of classic T7 promoter, new T7 Max promoter, and no template control sample. Samples were collected after an 8 -hour TxTl reaction. c: cell-free TxTl synthesis of GFP, T7 promoter (green bars) and T7Max promoter (blue bars), in house -made bacterial TxTl, with different ways of protecting linear DNA templates, and with commercially available myTXTL kit; end point fluorescence measured after 8 -hour reactions. For panels a, b and c each sample in triplicate, error bars are standard deviation. d example of Western Blot analysis of GFP translation, 8 -hour end point translation from linear dsDNA template in home-made TXTL without DNA protection reagents (samples represent conditions showed on panel c marked with red star). All TxTl reactions were incubated at 30°C