Team:Newcastle/13 July 2010
From 2010.igem.org
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- | ==Transformation== | + | ==LacI BioBrick Construction== |
+ | |||
+ | ===Aims=== | ||
+ | |||
+ | To use PCR to extract lacI (promoter, ribosome-binding site (RBS) & coding sequence (CDS)) from plasmid pMutin4 and ligate into vector pSB1AT3 in front of red fluorescent protein (RFP). | ||
+ | |||
+ | ===Transformation=== | ||
*''E. coli'' DH5alpha was transformed again with our plasmid. The [https://2010.igem.org/TeamNewcastleTransformation_of_E._coli#Protocol| transformation protocol] was followed. | *''E. coli'' DH5alpha was transformed again with our plasmid. The [https://2010.igem.org/TeamNewcastleTransformation_of_E._coli#Protocol| transformation protocol] was followed. | ||
- | ==Miniprep== | + | ===Miniprep=== |
*For this step the [https://2010.igem.org/Team:Newcastle/Qiagen_Minipreps| Qiagen Minipreps] protocol was followed. The 7 colonies that grew on the plates from yesterday were cultured ovenight on a shaking shelf at 37°C in universal tubes. | *For this step the [https://2010.igem.org/Team:Newcastle/Qiagen_Minipreps| Qiagen Minipreps] protocol was followed. The 7 colonies that grew on the plates from yesterday were cultured ovenight on a shaking shelf at 37°C in universal tubes. | ||
- | ==Restriction digests== | + | ===Restriction digests=== |
*The [https://2010.igem.org/TeamNewcastleRestriction_digests| restriction digest] protocol was followed for this step. | *The [https://2010.igem.org/TeamNewcastleRestriction_digests| restriction digest] protocol was followed for this step. | ||
{{Team:Newcastle/footer}} | {{Team:Newcastle/footer}} |
Revision as of 13:11, 6 August 2010
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Contents |
LacI BioBrick Construction
Aims
To use PCR to extract lacI (promoter, ribosome-binding site (RBS) & coding sequence (CDS)) from plasmid pMutin4 and ligate into vector pSB1AT3 in front of red fluorescent protein (RFP).
Transformation
- E. coli DH5alpha was transformed again with our plasmid. The transformation protocol was followed.
Miniprep
- For this step the Qiagen Minipreps protocol was followed. The 7 colonies that grew on the plates from yesterday were cultured ovenight on a shaking shelf at 37°C in universal tubes.
Restriction digests
- The restriction digest protocol was followed for this step.