Difference between revisions of "Cell-free chassis/Commercial E. coli S30"

(→‎Commercial ''E. coli'' S30)
(→‎Commercial ''E. coli'' S30)
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|<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
 
|<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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|<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 GFPmut3b synthesis (pmol 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. The error bars represent the standard deviation of the measurements.
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|<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 4. Graph of rate of GFPmut3b synthesis (pmol 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. The error bars represent the standard deviation of the measurements.
 
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|<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_Expression_Lifespan.JPG|thumb|200px|center|]] '''Graph 3. Graph of rate of GFPmut3b synthesis (pmol 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. The error bars represent the standard deviation of the measurements.
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|<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_Expression_Lifespan.JPG|thumb|200px|center|]] '''Graph 5. Graph of rate of GFPmut3b synthesis (pmol 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. The error bars represent the standard deviation of the measurements.
 
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|<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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|<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 6. Bar chart showing total GFPmut3b synthesized in 6 hours (pmol) 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. The error bars represent the standard deviation of the measurements.
 
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Revision as of 15:51, 25 October 2007

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

Temperature dependence
S30 Optimum Temp.JPG
Graph 1. Graph of GFPmut3b synthesized (pmol) 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.
DNA quantity dependence
 link
Product stability
 link
Peak time
S30 Peak Time.JPG
Graph 4. Graph of rate of GFPmut3b synthesis (pmol 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. The error bars represent the standard deviation of the measurements.
Expression lifespan
S30 Expression Lifespan.JPG
Graph 5. Graph of rate of GFPmut3b synthesis (pmol 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. The error bars represent the standard deviation of the measurements.
Expression capacity
S30 Expression Capacity.JPG
Graph 6. Bar chart showing total GFPmut3b synthesized in 6 hours (pmol) 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. The error bars represent the standard deviation of the measurements.


References

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