Team:Edinburgh/Notebook/Blue light producer
From 2010.igem.org
These notes correspond to the part of the project detailed here.
Blue Light Producer
5/7/2010
- LuxAB & LuxCDE operon agarose gel
- Lane1: About2.1.kbp
- Lane 2: The highest band- very faint- about 3.5kbp
13/7/2010
Empty edinbrick vector for control.
- LuxAB + edinbrick ligation.
28/7/2010
an update on LuxAB:
- The transformations of Edinbrick are fine but all the luxABs failed (grew blue, not white on Xgal and did not glow, as of 27/7)/
- Still have some digested LuxAB-ediI so re-did ligation and transformation.
Lane 1&15 | Lane 2 | Others |
Ladder | EdiI | LuxAB |
Lane 1 | Lane 2 +3 | Lanes 3-6 |
Ladder | EdiI | LuxAB |
29/7/2010
- The ligation had the same gel characteristics as previous luxAb-ediI ligations, e.x. it isn't visivle- not hopeful of this transformation working
4/8/2010
- made minipreps of blue (negative colonies)- double digest and ran in comparison with pure edinbrickI
- LacZ band dissapears but colonies still grow blue (WTF)
- Has bands equaling to luxAB and EdiI but there was no glowing
12/8/2010 update on LumP:
- still getting pink/red colonies from transformants
- single digests give a band of about 3kb
- double digests give a band of about 2.5 kb -highly confusing
- and also, no sign on RFP despite red/pink growth.
- Vector wa ssequenced and LumP is definately there.
Lanes 2-5: single digests of lumP Lane 6: double digest of lumP
26/8/2010
- Marker (Lane 1), minipreps 6-10 luxABlumP (lanes 2-6)
- single digests of LuxABlumP
7/9/2010
- minipreps of luxCDE: samples 1&4 usable
Lane 1 | Lane 2-6 | Lanes 7 |
Ladder | luxCDE00:1-5 minipreps | Ladder |