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Team:BCCS-Bristol/Notebook/Week 6
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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.]] | [[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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Revision as of 14:41, 9 October 2009
BCCS-Bristol
iGEM 2009
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.
