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| - | {{:Team:BCCS-Bristol/Header}}
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| - | ===Week 4===
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| - | * One more try to get Fiu protein on pSB1A3 backbone. NovaBlue transformed with ligation products of pSB1A3 and Fiu.
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| - | * Grew colonies for FhuA and OsmE midi preps.
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| - | * Carried out midi preps for FhuA and OsmE ( [FhuA]=378.9 ng/ul, [OsmE]=113.2 ng/ul).
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| - | * Transformed Nova Blue with biobrick part BBa_B0014 (double terminator)
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| - | * Culture colonies of BBa_B0014 transformants to get ready for mini prep.
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| - | * PCR FhuA and GFP for assembling together a quick and dirty fusion to be ready prior to BioScaffold arrival.
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| - | * Carried out colony growth (agar & liquid) for the AraC promoter (2 parts BBa_R0080 (lacking O2 region) and BBa_R0081 (the O2 region).
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| - | * Miniprepped DNA from cultures of BBa_R0080 and R0081.
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| - | * Attempted a 2-way ligation and of the BBa_R0080 and R0081 parts. The final product should be a 2-component biobrick on a plasmid in 5'-R0081-R0080-3' direction which can be transformed into cells.
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| - | * R0081 part was purified by gel extraction after being treated by double digest. R0080 and R0081 were ligated together and used to transform NovaBlue cells.
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| - | * First attempt to make the AraC promoter from the two parts failed.
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| - | * Fiu still not being ligated on ANY backbone.
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| - | [[Image:BCCS_Fhua_osme_midipreps.png|left|200px|frame|Standard 1kb NEB Ladder. Lanes 1-2 FhuA Midipreps, Lanes3-4 OsmE midipreps.]]
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| - | [[Image:BCCS_Fhua_gfp_pcr.png|center|200px|frame|Standard 1kb NEB Ladder. Lanes 1-2 FhuA PCR fragment for Quick and Dirty assembly Method, Lanes3-4 GFP PCR fragments for quick and dirty assembly method.]]
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| - | [[Image:BCCS_R0080_r0081_bw.png|center|500px|frame|The figure indicates the presence of the AraC promoter subunits. Lanes 1-6 show the unsuccesful ligation of the Fiu Carrier protein. Lanes 7-9 illustrate Double Digest with XbaI and SpeI,Single Digest with XbaI and Uncut versions of the plasmid carrying R0080. Similarly lanes 10-12 show the same for R0081. The promoter fragments are 149bp and 183bp respectively and do not appear on the gel but the presence is
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| - | illustrated by the band shift between the Double and Single Digests.]]
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| - | {{:Team:BCCS-Bristol/NotebookHeader}}
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