Team:Newcastle/15 July 2010

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=LacI BioBrick Construction=
 
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==Aims==
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=Research=
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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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==Materials==
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We initiated our research on subtilin quorum sensing system which would provide a coordinated cell response.
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*[[Team:Newcastle/8_July_2010|Original transformants
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*[[Team:Newcastle/14_July_2010|Second round of transformants]]
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*[[Team:Newcastle/12_July_2010|Plasmid extracts]]
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*Restriction enzymes ''Xba''1 aand ''Pst''1
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==Protocol==
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#Klein C, Kaletta C and Entian KD. (1993). "Biosynthesis of the lantibiotic subtilin is regulated by a histidine kinase/response regulator system". ''Applied and environmental microbiology''. 59(1). 296-303.
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*Single (''Xba''1) and Double (''Xba''1 aand ''Pst''1) digest of Plasmid extracts
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#Heinzmann S, Entian KD and Stein T. (2006). "Engineering Bacillus subtilis ATCC 6633 for improved production of the lantibiotic subtilin". ''Applied microbiology and biotechnology''. 69(5). 532-6.  
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*Run digests on a gel
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*Overnight culture of all transformants
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==Results==
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'''Figure 1.'''
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This gel is unclear in precisely what is present in the samples it can clearly be seen that there are in fact two bands in the single digest lanes (where there should only be one) and three bands in the double digest lane (where there should only be two).
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==Conclusion==
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It is our current belief that the [[Team:Newcastle/13_July_2010|plasmid extraction]] may have been performed incorrectly (there was one minor mistake during the procedure), so to this end an overnight culture of the original transformant colonies, as well as second round of transformants, will be performed, ready for plasmid extraction tomorrow.  
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{{Team:Newcastle/footer}}
{{Team:Newcastle/footer}}

Latest revision as of 20:19, 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. Klein C, Kaletta C and Entian KD. (1993). "Biosynthesis of the lantibiotic subtilin is regulated by a histidine kinase/response regulator system". Applied and environmental microbiology. 59(1). 296-303.
  2. Heinzmann S, Entian KD and Stein T. (2006). "Engineering Bacillus subtilis ATCC 6633 for improved production of the lantibiotic subtilin". Applied microbiology and biotechnology. 69(5). 532-6.
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