Difference between revisions of "Cell-free chassis/Commercial E. coli T7 S30"
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=Commercial ''E. coli'' T7 S30= | =Commercial ''E. coli'' T7 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> | |
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| − | | | + | |<center>[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems/Commercial_E.coli_T7_S30 <font face=georgia color=#000066 size=4>Chassis description</font>]</center> || The Commercial ''E. coli'' T7 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 Commercial ''E. coli'' T7 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_E7104 BBa_E7104] has been used to characterize this cell-free chassis | The simple constitutive gene expression device [https://parts.igem.org/wiki/index.php/Part:BBa_E7104 BBa_E7104] has been used to characterize this cell-free chassis | ||
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| + | |<center>[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems/Commercial_E.coli_T7_S30/Optimum_Temp <font face=georgia color=#000066 size=4>Temperature dependence</font>]</center> || [[Image:T7_Optimum_Temp.JPG|thumb|200px|center|]] '''Graph 1. Graph of fluorescence (au) against time (minutes) at 4oC (blue), 25oC (red) and 37oC (green).''' The simple constitutive gene expression device [https://parts.igem.org/wiki/index.php/Part:BBa_E7104 BBa_E7104] has been used. Each of the three 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 OpenWetWare for the detailed protocol and raw data of the experiment. | ||
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| + | |<center>[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems/Commercial_E.coli_T7_S30/Optimum_DNA <font face=georgia color=#000066 size=4>Optimum amount of DNA</font>]</center> || link | ||
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| + | |<center>[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems/Commercial_E.coli_T7_S30/Product_Stability <font face=georgia color=#000066 size=4>Product stability</font>]</center> || link | ||
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| + | |<center>[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems/Commercial_E.coli_T7_S30/Peak_Time <font face=georgia color=#000066 size=4>Peak time</font>]</center> || [[Image:T7_Peak_Time.JPG|thumb|200px|center|]]'''Graph 2. Graph of rate of GFP synthesis (mol per min) against time (minutes) at 4oC (blue), 25oC (red) and 37oC (green).''' The simple constitutive gene expression device [https://parts.igem.org/wiki/index.php/Part:BBa_E7104 BBa_E7104] has been used. Each of the three 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 OpenWetWare for the detailed protocol and raw data of the experiment. | ||
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| + | |<center>[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems/Commercial_E.coli_T7_S30/Expression_Lifespan <font face=georgia color=#000066 size=4>Expression lifespan</font>]</center> || [[Image:T7_Peak_Time.JPG|thumb|200px|center|]] '''Graph 2. Graph of rate of GFP synthesis (mol per min) against time (minutes) at 4oC (blue), 25oC (red) and 37oC (green).''' The simple constitutive gene expression device [https://parts.igem.org/wiki/index.php/Part:BBa_E7104 BBa_E7104] has been used. Each of the three 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 OpenWetWare for the detailed protocol and raw data of the experiment. | ||
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| + | |<center>[https://parts.igem.org/wiki/index.php/Chassis/Cell-Free_Systems/Commercial_E.coli_T7_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 (blue), 25oC (red) and 37oC (green).''' The simple constitutive gene expression device [https://parts.igem.org/wiki/index.php/Part:BBa_E7104 BBa_E7104] has been used. Each of the three coloured bars show average measurements based on three replicates of the experiment. The error bars represent the standard deviation of the measurements. This bar chart chart is derived from Graph 1, with addition data from our GFP calibration curve. Please visit our OpenWetWare for the detailed protocol and raw data of the experiment. | ||
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==References== | ==References== | ||
Please refer to the [http://www.promega.com/tbs/tb219/tb219.pdf Promega cell extract user guide] for detailed information about the Commercial ''E. coli'' T7 S30 extract. | Please refer to the [http://www.promega.com/tbs/tb219/tb219.pdf Promega cell extract user guide] for detailed information about the Commercial ''E. coli'' T7 S30 extract. | ||
Revision as of 10:07, 25 October 2007
Commercial E. coli T7 S30
| Cell-Free Systems (back to Intro) | Introduction |
| The Commercial E. coli T7 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_E7104 has been used to characterize this cell-free chassis | |
| Graph 1. Graph of fluorescence (au) against time (minutes) at 4oC (blue), 25oC (red) and 37oC (green). The simple constitutive gene expression device BBa_E7104 has been used. Each of the three 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 OpenWetWare for the detailed protocol and raw data of the experiment. | |
| link | |
| link | |
| Graph 2. Graph of rate of GFP synthesis (mol per min) against time (minutes) at 4oC (blue), 25oC (red) and 37oC (green). The simple constitutive gene expression device BBa_E7104 has been used. Each of the three 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 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 (blue), 25oC (red) and 37oC (green). The simple constitutive gene expression device BBa_E7104 has been used. Each of the three 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 OpenWetWare for the detailed protocol and raw data of the experiment. | |
| Graph 3. Bar chart showing total GFP synthesized in 6 hours (mol) at 4oC (blue), 25oC (red) and 37oC (green). The simple constitutive gene expression device BBa_E7104 has been used. Each of the three coloured bars show average measurements based on three replicates of the experiment. The error bars represent the standard deviation of the measurements. This bar chart chart is derived from Graph 1, with addition data from our GFP calibration curve. Please visit our OpenWetWare for the detailed protocol and raw data of the experiment. |
References
Please refer to the [http://www.promega.com/tbs/tb219/tb219.pdf Promega cell extract user guide] for detailed information about the Commercial E. coli T7 S30 extract.