Team:Imperial College/Biobricks

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Revision as of 22:38, 29 October 2008

Biobrick Characterisation in B.subtilis
This year Imperial College has added 45 new B.subtilis specific parts to the registry. In addition we sought to characterise some of these parts, including the constitutive promoter and ribosome binding site pveg-spoVG. The testing constructs we developed for this are shown below:

Summary of Data

The promoter and RBS combinations were characterised by measuring the expression of fluorescent proteins in B.subtilis. Cultures of B.subtilis transformed with the test constructs and non-transformed B.subtilis (control) were grown to the mid-log phase. Fluorescence and O.D.₆₀₀ were both measured using a plate reader to generate fluorescence levels of the various samples. To make this data more generic, the fluorescence data were normalized based on cell number using the O.D.₆₀₀ and a calibration curve of O.D.₆₀₀ vs colony forming units, as explained in the diagram below:

This calibration curve allowed us to produce the final data as shown below. This clearly shows that B.subtilis transformed with BioBricks GFP and RFP are fluorescing at the corresponding excitation and emission wavelengths.

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 <br>
 {{Imperial/Box2|Biobrick Characterisation in ''B.subtilis''|
-This year Imperial College has added 45 new ''B.subtilis'' specific parts to the registry. In addition we sort to characterise some of these parts, including the constitutive promoter and ribosome binding site '''pveg-spoVG'''. The testing constructs we developed for this are shown below:
+This year Imperial College has added 45 new ''B.subtilis'' specific parts to the registry. In addition we sought to characterise some of these parts, including the constitutive promoter and ribosome binding site '''pveg-spoVG'''. The testing constructs we developed for this are shown below:
 [[Image:Biobricks Imperial.PNG|center|500px]]
 }}
-{{Imperial/Box1|Summary of Data|
+{{Imperial/Box1|Summary of Data|The promoter and RBS combinations were characterised by measuring the expression of fluorescent proteins in ''B.subtilis''. Cultures of ''B.subtilis'' transformed with the test constructs and non-transformed ''B.subtilis'' (control) were grown to the mid-log phase. Fluorescence and O.D.<sub>600</sub> were both measured using a plate reader to generate fluorescence levels of the various samples. To make this data more generic, the fluorescence data were normalized based on cell number using the O.D.<sub>600</sub> and a '''calibration curve of O.D.<sub>600</sub> vs colony forming units''', as explained in the diagram below:
-The promoter and RBS combinations were characterised by measuring the expression of fluorescent proteins in ''B.subtilis''. Cultures of ''B.subtilis'' transformed with the test constructs and non-transformed ''B.subtilis'' (control) were grown to the mid-log phase. Fluorescence and O.D.<sub>600</sub> were both measured using a plate reader to generate fluorescence levels of the various samples. To make this data more generic the fluorescence data was normalized based on cell number using the O.D.<sub>600</sub> and a '''calibration curve of O.D.<sub>600</sub> vs colony forming units''', as explained in the diagram below:
 <br>
 [[Image:Calibration Picture.PNG|600px|center]]
 <br>
-This calibration curve allowed us to produce the final data as shown below. This clearly shows ''B.subtilis'' transformed with biobricks GFP and RFP are fluorescing at the corresponding excitation and emission wavelengths.
+This calibration curve allowed us to produce the final data as shown below. This clearly shows that ''B.subtilis'' transformed with BioBricks GFP and RFP are fluorescing at the corresponding excitation and emission wavelengths.
 <br>
 [[Image:Fluorescence B.subtilis.PNG|400px|center]]