Difference between revisions of "IGEM:IMPERIAL/2008/New/Protocols"

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= '''Protocols''' =
 
  
{| cellpadding="5" style="background:#66aadd; border: 4px solid #66aadd; color:black" align="center" width=100%
 
!width="100%"|
 
  
<font size=4px><font color=white>'''1. Transformation Protocols for integration vectors in ''Bacillus subtilis'''''
+
{{Imperial/Box2|Experiments & Protocols|
 +
Below you will find a comprehensive list of all the protocols we used during our wet lab work. We spent some time toward the start of the project sourcing and updating effective protocols for ''B. subtilis'' manipulation explicitly, as other teams have previously had difficulty in this area.
 +
 
 +
These protocols worked for us, so any future teams looking to use ''subtilis'' could save quite a bit of time by looking here before diving into the literature!}}
 +
 
 +
 
 +
{| cellpadding="1" style="background:#66aadd; border:1px solid #66aadd; color:black" align="center" width=50%
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|width="100%"| <font size=4px color=white>'''1. Transformation Protocols for integration vectors in ''Bacillus subtilis'''''</font>
 
|-
 
|-
 
|style="background:#FFFFFF"|  
 
|style="background:#FFFFFF"|  
 
*[[/Transformation_protocol_2 | Protocol 1]]
 
*[[/Transformation_protocol_2 | Protocol 1]]
 
*[[/Transformation_protocol_4 | Protocol 2]]
 
*[[/Transformation_protocol_4 | Protocol 2]]
 
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|-
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|style="background:#66aadd"| <font size=4px color=white>'''2. Transformation Protocol for linear DNA in ''Bacillus subtilis'''''</font>
 
|-
 
|-
cellpadding="5" style="background:#66aadd; border: 4px solid #66aadd; color:black" align="center" width=100%
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|style="background:#FFFFFF"|
!width="100%"|
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*Protocol
 
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|-
<font size=4px><font color=white>'''2. Transformation Protocol for linear DNA in ''Bacillus subtilis'''''
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|style="background:#66aadd"| <font size=4px color=white>'''3. Transformation Protocol ''E.coli'''''</font>
 
 
 
|-
 
|-
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|style="background:#FFFFFF"|
!width="100%"|
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*[[/XL1-Blue_preparation | Preparation of XL1-blue electrocompetent cells]]
 
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|-
<font size=4px><font color=white>'''3. Transformation Protocol ''E.coli'''''
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|style="background:#66aadd"| <font size=4px color=white>'''4. Motility Analysis'''</font>
 
 
 
|-
 
|-
 
|style="background:#FFFFFF"|  
 
|style="background:#FFFFFF"|  
*[[/XL1-Blue_preparation | Preparation of XL1-blue electrocompetent cells]]
 
 
 
|}
 
 
 
 
 
 
 
 
4. Motility Analysis
 
 
*[[/Protocols#Motility_Experiments | Motility Assays]]
 
*[[/Protocols#Motility_Experiments | Motility Assays]]
5. PCR
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|-
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|style="background:#66aadd"| <font size=4px color=white>'''5. PCR'''</font>
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|-
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|style="background:#FFFFFF"|
 
*[[/PCR| PCR]]
 
*[[/PCR| PCR]]
 
*[[/Colony_PCR| Single Colony PCR]]
 
*[[/Colony_PCR| Single Colony PCR]]
6. Calibration Curves<br>
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|-
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|style="background:#66aadd"| <font size=4px color=white>'''6. Calibration Curves'''</font>
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|-
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|style="background:#FFFFFF"|
 
*[[/Calibration-Curves| Calibration Curves ]]
 
*[[/Calibration-Curves| Calibration Curves ]]
 
*[[/Growth| O.D.<sub>600</sub> vs cell number]]
 
*[[/Growth| O.D.<sub>600</sub> vs cell number]]
 
*[[/Fluorescence| Fluorescence vs GFPmut3b]]
 
*[[/Fluorescence| Fluorescence vs GFPmut3b]]
7. Promoter-RBS Testing Protocols<br>
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|-
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|style="background:#66aadd"| <font size=4px color=white>'''7. Promoter-RBS Testing Protocols'''</font>
 +
|-
 +
|style="background:#FFFFFF"|
 
*[[/Constitutive | Constitutive Promoters and RBS]]  
 
*[[/Constitutive | Constitutive Promoters and RBS]]  
 
*[[/Inducible | pHyperspank Promoter and RBS]]
 
*[[/Inducible | pHyperspank Promoter and RBS]]
 
*[[/Light_Inducible | Light Inducible Promoters and RBS]]
 
*[[/Light_Inducible | Light Inducible Promoters and RBS]]
8. Biomaterial Protocols
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|-
 
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|style="background:#66aadd"| <font size=4px color=white>'''8. Biomaterial Protocols'''</font>
 
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|-
 
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|style="background:#FFFFFF"|
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*Protocol
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|}
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<br clear="all"><br><hr>
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{{Imperial/EndPage|Cloning_Strategy|Major_Results}}

Latest revision as of 04:20, 30 September 2008

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       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Project">Project Specifications</a>
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Chassis_1">Why B. subtilis?</a>
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Chassis_2">B. subtilis: Benefits vs Challenges</a>
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       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Protocols">Experiments & Protocols</a>
       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Major_Results">Experimental Results</a>
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       <a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Appendices">Appendices - Code etc.</a>
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</td><td align="center" width="17%" valign="bottom"><ul id="sddm"><a href="http://2008.igem.org/Team:Imperial_College/Notebook"> Notebook </a></ul> </td><td align="center" width="17%" valign="bottom"><ul id="sddm"><a href="http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Team"> Our Team </a></ul> </td></tr></table></html>

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Experiments & Protocols

Below you will find a comprehensive list of all the protocols we used during our wet lab work. We spent some time toward the start of the project sourcing and updating effective protocols for B. subtilis manipulation explicitly, as other teams have previously had difficulty in this area.

These protocols worked for us, so any future teams looking to use subtilis could save quite a bit of time by looking here before diving into the literature!



1. Transformation Protocols for integration vectors in Bacillus subtilis
2. Transformation Protocol for linear DNA in Bacillus subtilis
  • Protocol
3. Transformation Protocol E.coli
4. Motility Analysis
5. PCR
6. Calibration Curves
7. Promoter-RBS Testing Protocols
8. Biomaterial Protocols
  • Protocol




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