Team:BCCS-Bristol/Notebook/Week 4

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~~{{:Team:BCCS-Bristol/Header}}~~

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~~{{:Team:BCCS-Bristol/NotebookHeader}}~~

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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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-{{:Team:BCCS-Bristol/Header}}
-{{:Team:BCCS-Bristol/NotebookHeader}}
-===Week 4===
-* One more try to get Fiu protein on pSB1A3 backbone. NovaBlue transformed with ligation products of pSB1A3 and Fiu.
-* Grew colonies for FhuA and OsmE midi preps.
-* Carried out midi preps for FhuA and OsmE ( [FhuA]=378.9 ng/ul, [OsmE]=113.2 ng/ul).
-* Transformed Nova Blue with biobrick part BBa_B0014 (double terminator)
-* Culture colonies of BBa_B0014 transformants to get ready for mini prep.
-* PCR FhuA and GFP for assembling together a quick and dirty fusion to be ready prior to BioScaffold arrival.
-* Carried out colony growth (agar & liquid) for the AraC promoter (2 parts BBa_R0080 (lacking O2 region) and BBa_R0081 (the O2 region).
-* Miniprepped DNA from cultures of BBa_R0080 and R0081.
-* 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.
-* 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.
-* First attempt to make the AraC promoter from the two parts failed.
-* Fiu still not being ligated on ANY backbone.
-[[Image:BCCS_Fhua_osme_midipreps.png|left|200px|frame|Standard 1kb NEB Ladder. Lanes 1-2 FhuA Midipreps, Lanes3-4 OsmE midipreps.]]
-[[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.]]

Team:BCCS-Bristol/Notebook/Week 4

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Latest revision as of 23:48, 20 October 2009