Team:BCCS-Bristol/Notebook/Week 6

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{{:Team:BCCS-Bristol/Header}}
 
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===Week 6===
 
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* XL1-Blue transformations with ligations 1 and 2 from Week 5.
 
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* Restriction digests for FhuA and OsmE to prepare their insertion downstream of the AraC-RBS costruct. Also GFP cut to fuse it with BOO14 terminator.
 
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* Ligation set up between GFP and B0014 terminator (on pSB1AK3) and used to transform XL1-Blue.
 
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* Transformations with AraC-RBS and pQE31-GFP didn't work. Repeated with NovaBlue cells.
 
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* AraC-RBS transformations worked very well in contrast with pQE31-GFP and GFP-terminator that didn't work again.
 
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* Repeat ligations between pQE31-GFP and GFP-terminator and transform XL1-Blue.
 
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* Minipreps and Restr. Digests for AraC-RBS constructs.
 
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* pQE31-GFP and GFP-terminator transformations failed.
 
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* Ligations set up:
 
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1. pQE31+GFP
 
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2. AraC-RBS + FhuA
 
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3. AraC-RBS + OsmE
 
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4. GFP-terminator
 
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[[Image:BCCS_Promoter_rbs_presence.jpg|center|100px|frame|1kb NEB Standard Ladder. Lanes run in triplets featuring Double Digest (XbaI/SpeI), Single Digest(XbaI) and uncut versions of DNA plasmid carrying the AraC-RBS ligation product. The product is seen as a single band at the bottom of the Double Digest lanes. The top band is the pSB1A2 Carrier plasmid.]]
 
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{{:Team:BCCS-Bristol/NotebookHeader}}
 

Latest revision as of 23:48, 20 October 2009