Team:BCCS-Bristol/Notebook/Week 6

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BCCS-Bristol
iGEM 2009


Week 6

  • XL1-Blue transformations with ligations 1 and 2 from Week 5.
  • 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.
  • Ligation set up between GFP and B0014 terminator (on pSB1AK3) and used to transform XL1-Blue.
  • Transformations with AraC-RBS and pQE31-GFP didn't work. Repeated with NovaBlue cells.
  • AraC-RBS transformations worked very well in contrast with pQE31-GFP and GFP-terminator that didn't work again.
  • Repeat ligations between pQE31-GFP and GFP-terminator and transform XL1-Blue.
  • Minipreps and Restr. Digests for AraC-RBS constructs.
  • pQE31-GFP and GFP-terminator transformations failed.
  • Ligations set up: 
1. pQE31+GFP
2. AraC-RBS + FhuA
3. AraC-RBS + OsmE
4. GFP-terminator


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.


Week Zero Week 1 Week 2 Week 3 Week 4 Week 5 Week 6 Week 7 Week 8 Week 9 Week 10 Week 11 Week 12
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