Cell-free chassis/Commercial E. coli S30

Commercial E. coli S30

Cell-Free Systems (back to Intro)	Introduction
Chassis description	The Commercial E. coli S30 extract was purchased from Promega. It was prepared by modifications of the method described by Zubay et al (1980). E. coli strain B deficient in OmpT endoproteinase and lon protease activity was used. The simple constitutive gene expression device BBa_I13522 has been used to characterize this cell-free chassis
Optimum temperature	Graph 1. Graph of fluorescence (au) against time (minutes) at 4oC (dark blue), 15oC (pink), 20oC (yellow) and 30oC (light blue). The simple constitutive gene expression device BBa_I13522 has been used. Each of the four coloured lines show average measurements based on three replicates of the experiment. The error bars represent the standard deviation of the measurements. Please visit our [http://openwetware.org/wiki/IGEM:IMPERIAL/2007/Wet_Lab/Protocols/CBD2.2 OpenWetWare] for the detailed protocol and raw data of the experiment.
Peak time	Graph 2. Graph of rate of GFP synthesis (mol per min) against time (minutes) at 4oC (dark blue), 15oC (pink), 20oC (yellow) and 30oC (light blue). The simple constitutive gene expression device BBa_I13522 has been used. Each of the four coloured lines show average measurements based on three replicates of the experiment. This graph is derived from Graph 1, with addition data from our GFP calibration curve. Please visit our [http://openwetware.org/wiki/IGEM:IMPERIAL/2007/Wet_Lab/Protocols/CBD2.2 OpenWetWare] for the detailed protocol and raw data of the experiment.
Product stability	link
Expression capacity	Graph 3. Bar chart showing total GFP synthesized in 6 hours (mol) at 4oC (dark blue), 15oC (pink), 20oC (yellow) and 30oC (light blue). The simple constitutive gene expression device BBa_I13522 has been used. Each of the four coloured bars show average measurements based on three replicates of the experiment. This graph is derived from Graph 1, with addition data from our GFP calibration curve. Please visit our [http://openwetware.org/wiki/IGEM:IMPERIAL/2007/Wet_Lab/Protocols/CBD2.2 OpenWetWare] for the detailed protocol and raw data of the experiment.
Expression lifespan	Graph 2. Graph of rate of GFP synthesis (mol per min) against time (minutes) at 4oC (dark blue), 15oC (pink), 20oC (yellow) and 30oC (light blue). The simple constitutive gene expression device BBa_I13522 has been used. Each of the four coloured lines show average measurements based on three replicates of the experiment. This graph is derived from Graph 1, with addition data from our GFP calibration curve. Please visit our [http://openwetware.org/wiki/IGEM:IMPERIAL/2007/Wet_Lab/Protocols/CBD2.2 OpenWetWare] for the detailed protocol and raw data of the experiment.

References

Please refer to the Promega cell extract user guide for detailed information about the Commercial E. coli S30 extract.