Team:DTU-Denmark/Notebook/24 August 2013

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24 August 2013

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Contents

Lab 208


Main purpose


  • fix problem with gels
  • run gels

Who was in the lab


Kristian, Henrike

Procedure


Running and loading gels.

Results


Gels

  • 1 kb
  • colony PCR NirG
  • constitutive reference promoter, GC buffer
  • constitutive reference promoter, GC buffer + 3% DMSO
  • constitutive reference promoter, HF buffer
  • constitutive reference promoter, HF buffer + 3% DMSO
  • 1 kb

2013-08-24 nircol ref const.jpg

Gel could not be read and will be redone.

  • 1 kb
  • colony PCR cycAX, colony 2
  • colony PCR cycAX, colony 3
  • colony PCR cycAX, colony 4
  • colony PCR cycAX, colony 6
  • colony PCR cycAX, colony 7
  • colony PCR cycAX, colony 8
  • colony PCR cycAX, colony 9
  • constitutive reference promoter, GC buffer
  • constitutive reference promoter, GC buffer + 3% DMSO
  • constitutive reference promoter, HF buffer
  • constitutive reference promoter, HF buffer + 3% DMSO
  • 1 kb

2013-08-24 cyccol refconst.jpg

The insert is confirmed for the colonies 3, 4 and 8.

Gel optimization test

Made two gels with TAE buffer to run on the two different power supplies and used two different loading dyes to try and resolve the problem with the bad gels we've been having.

Loading dyes: Agarose Blue was made by us, GelPilot is supplied by Qiagen. The const ref samples were mixed with agarose blue before, so we cannot test the other loading dye on those.

Run on the old power supply (the grey one):

  • 1 kb
  • cycAX col 3, agarose blue
  • cycAX col 4, GelPilot
  • cycAX col 8, GelPilot
  • cycAX col 2, agarose blue
  • cycAX col 6, agarose blue
  • cycAX col 7, GelPilot
  • cycAX col 9, GelPilot
  • constitutive reference promoter, GC buffer
  • constitutive reference promoter, GC buffer + 3% DMSO
  • constitutive reference promoter, HF buffer
  • constitutive reference promoter, HF buffer + 3% DMSO
  • NirG col, agarose blue
  • NirG col, GelPilot
  • HAO col, agarose blue
  • HAO col, GelPilot
  • 1 kb

2013-08-24 check big.jpg

Run on the new power supply (the green one):

  • 1 kb
  • cycAX col 3, agarose blue
  • cycAX col 4, GelPilot
  • cycAX col 8, GelPilot
  • cycAX col 2, agarose blue
  • cycAX col 7, GelPilot
  • constitutive reference promoter, GC buffer
  • constitutive reference promoter, GC buffer + 3% DMSO
  • constitutive reference promoter, HF buffer
  • cycAX col 3, GelPilot
  • NirG col, agarose blue
  • NirG col, GelPilot
  • HAO col, agarose blue
  • HAO col, GelPilot
  • 1 kb

2013-08-24 check small.jpg

Conclusion


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