Team:Newcastle/14 July 2010

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=LacI BioBrick Construction=
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=Research=
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==Aims==
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We initiated our research on subtilin quorum sensing system which would provide a coordinated cell response.
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*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).
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*To see whether the LacI insert is in the pSB1AT3 plasmid (same as yesterday).  
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==Materials and Protocol==
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#Kleerebezem M, Bongers R, Rutten G, Vos WM de and Kuipers OP. (2004). "Autoregulation of subtilin biosynthesis in Bacillus subtilis: the role of the spa-box in subtilin-responsive promoters". ''Peptides''. 25(9). 1415-24.  
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*Performed a gel electrophoresis  with remaining PCR products + Controls with same restriction digests.  
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#Klein C, Kaletta C, Schnell N and Entian KD. (1992). "Analysis of genes involved in biosynthesis of the lantibiotic subtilin". ''Applied and environmental microbiology''. 58(1). 132-42.
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====Transformation====
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*''E. coli'' DH5alpha was transformed again with our plasmid. The [https://2010.igem.org/TeamNewcastleTransformation_of_E._coli#Protocol| transformation protocol] was followed.  
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====Miniprep====
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*For this step the [[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.
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====Restriction digests====
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*The [https://2010.igem.org/TeamNewcastleRestriction_digests| restriction digest] protocol was followed for this step.
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(we followed Wendy's cards/Phil's lab-book)
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{{Team:Newcastle/footer}}
{{Team:Newcastle/footer}}

Latest revision as of 20:15, 25 October 2010

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Research

We initiated our research on subtilin quorum sensing system which would provide a coordinated cell response.

  1. Kleerebezem M, Bongers R, Rutten G, Vos WM de and Kuipers OP. (2004). "Autoregulation of subtilin biosynthesis in Bacillus subtilis: the role of the spa-box in subtilin-responsive promoters". Peptides. 25(9). 1415-24.
  2. Klein C, Kaletta C, Schnell N and Entian KD. (1992). "Analysis of genes involved in biosynthesis of the lantibiotic subtilin". Applied and environmental microbiology. 58(1). 132-42.
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