Imperial College/21 August 2008
From 2008.igem.org
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* Continued work on modelling the growth curve of bacteria using ODEs | * Continued work on modelling the growth curve of bacteria using ODEs | ||
- | == Wet Lab== | + | == Wet Lab== |
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+ | ===Cloning=== | ||
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+ | * Double digest (''EcoRI'' and ''SpeI'') of all registry sequences performed and insert sizes checked on gel | ||
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===''B.subtilis''=== | ===''B.subtilis''=== | ||
* The ''B.subtilis'' was successfully transformed using transformation of protocol 2, [http://openwetware.org/wiki/IGEM:IMPERIAL/2008/Prototype/Wetlab/Transformation_protocol_4#Transformation_Protocol_2| Click link here for protocol]. However, colonies were seen on the plates were no DNA was used for transformation. The most likely cayse of this is contamination whilst carrying out the electroporation. To check that the LB agar plates contained streptinomycin we picked a transformed colony and streaked it out on a freshly made LB agar plate containing streptinmycin. | * The ''B.subtilis'' was successfully transformed using transformation of protocol 2, [http://openwetware.org/wiki/IGEM:IMPERIAL/2008/Prototype/Wetlab/Transformation_protocol_4#Transformation_Protocol_2| Click link here for protocol]. However, colonies were seen on the plates were no DNA was used for transformation. The most likely cayse of this is contamination whilst carrying out the electroporation. To check that the LB agar plates contained streptinomycin we picked a transformed colony and streaked it out on a freshly made LB agar plate containing streptinmycin. | ||
*In addition the transformation protocol 1 was carried out, [http://openwetware.org/wiki/IGEM:IMPERIAL/2008/Prototype/Wetlab/Transformation_protocol_2| click here for a link to the protocol]. The principle of this protocol is that a culture is grown to a high O.D.<sub>600</sub>, this induces the stress pathways in ''B.subtilis'' and induces natural competence. These high cultures are diluted and then incubated with suitable concentrations of DNA. Transformed ''B.subtilis'' can then be plated out and grown on antibiotic resistant plates. Today ''B.subtilis'' were grown to an O.D.<sub>600</sub> of 2.0 and incubated with 40ng to 400ng of pDR110 DNA. | *In addition the transformation protocol 1 was carried out, [http://openwetware.org/wiki/IGEM:IMPERIAL/2008/Prototype/Wetlab/Transformation_protocol_2| click here for a link to the protocol]. The principle of this protocol is that a culture is grown to a high O.D.<sub>600</sub>, this induces the stress pathways in ''B.subtilis'' and induces natural competence. These high cultures are diluted and then incubated with suitable concentrations of DNA. Transformed ''B.subtilis'' can then be plated out and grown on antibiotic resistant plates. Today ''B.subtilis'' were grown to an O.D.<sub>600</sub> of 2.0 and incubated with 40ng to 400ng of pDR110 DNA. |
Revision as of 19:11, 25 August 2008
21 August 2008Dry Lab
Wet LabCloning
B.subtilis
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