Difference between revisions of "Cell-free chassis/Commercial E. coli S30"
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|[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems <font face=georgia color=#cc0033 size=4>Cell-Free Systems (back to Intro)</font>] || <font face=georgia size=4>Introduction</font> | |[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems <font face=georgia color=#cc0033 size=4>Cell-Free Systems (back to Intro)</font>] || <font face=georgia size=4>Introduction</font> | ||
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| − | |[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems/Commercial_E.coli_S30 <font face=georgia color=#000066 size=4>Chassis description</font>] || 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. | + | |<center>[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems/Commercial_E.coli_S30 <font face=georgia color=#000066 size=4>Chassis description</font>]</center> || 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. |
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The simple constitutive gene expression device [https://parts.igem.org/wiki/index.php/Part:BBa_I13522 BBa_I13522] has been used to characterize this cell-free chassis | The simple constitutive gene expression device [https://parts.igem.org/wiki/index.php/Part:BBa_I13522 BBa_I13522] has been used to characterize this cell-free chassis | ||
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| − | |[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems/Commercial_E.coli_S30/Optimum_Temp <font face=georgia color=#000066 size=4>Optimum temperature</font>] || [[Image:S30_Optimum_Temp.JPG|thumb|200px|center|]] '''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 [https://parts.igem.org/wiki/index.php/Part:BBa_I13522 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. | + | |<center>[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems/Commercial_E.coli_S30/Optimum_Temp <font face=georgia color=#000066 size=4>Optimum temperature</font>]</center> || [[Image:S30_Optimum_Temp.JPG|thumb|200px|center|]] '''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 [https://parts.igem.org/wiki/index.php/Part:BBa_I13522 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. |
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| − | |[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems/Commercial_E.coli_S30/Peak_Time <font face=georgia color=#000066 size=4>Peak time</font>] || [[Image:S30_Peak_Time.JPG|thumb|200px|center|]]'''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 [https://parts.igem.org/wiki/index.php/Part:BBa_I13522 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. | + | |<center>[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems/Commercial_E.coli_S30/Peak_Time <font face=georgia color=#000066 size=4>Peak time</font>]</center> || [[Image:S30_Peak_Time.JPG|thumb|200px|center|]]'''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 [https://parts.igem.org/wiki/index.php/Part:BBa_I13522 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. |
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| − | |[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems/Commercial_E.coli_S30/Product_Stability <font face=georgia color=#000066 size=4>Product stability</font>] || link | + | |<center>[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems/Commercial_E.coli_S30/Product_Stability <font face=georgia color=#000066 size=4>Product stability</font>]</center> || link |
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| − | |[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems/Commercial_E.coli_S30/Expression_Capacity <font face=georgia color=#000066 size=4>Expression capacity</font>] || [[Image:S30_Expression_Capacity.JPG|thumb|200px|center|]] '''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 [https://parts.igem.org/wiki/index.php/Part:BBa_I13522 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. | + | |<center>[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems/Commercial_E.coli_S30/Expression_Capacity <font face=georgia color=#000066 size=4>Expression capacity</font>]</center> || [[Image:S30_Expression_Capacity.JPG|thumb|200px|center|]] '''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 [https://parts.igem.org/wiki/index.php/Part:BBa_I13522 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. |
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| − | |[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems/Commercial_E.coli_S30/Expression_Lifespan <font face=georgia color=#000066 size=4>Expression lifespan</font>] || [[Image:S30_Peak_Time.JPG|thumb|200px|center|]] '''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 [https://parts.igem.org/wiki/index.php/Part:BBa_I13522 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. | + | |<center>[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems/Commercial_E.coli_S30/Expression_Lifespan <font face=georgia color=#000066 size=4>Expression lifespan</font>]</center> || [[Image:S30_Peak_Time.JPG|thumb|200px|center|]] '''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 [https://parts.igem.org/wiki/index.php/Part:BBa_I13522 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. |
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Revision as of 09:52, 25 October 2007
Commercial E. coli S30
| Cell-Free Systems (back to Intro) | Introduction |
| 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 | |
| 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. | |
| 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. | |
| link | |
| 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. | |
| 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.