Team:Edinburgh/Notebook/Blue light producer

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Revision as of 06:45, 27 October 2010 by JRWK (Talk | contribs)





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

150.JPG

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.

154.JPG

Lane 1&15 Lane 2 Others
Ladder EdiI LuxAB

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Lane 1 Lane 2 +3 Lanes 3-6
Ladder EdiI LuxAB

29/7/2010

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  • 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

162.jpg

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.

160.jpg

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)

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  • single digests of LuxABlumP

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7/9/2010

  • minipreps of luxCDE: samples 1&4 usable
Lane 1 Lane 2-6 Lanes 7
Ladder luxCDE00:1-5 minipreps Ladder

140.JPG