Team:DTU-Denmark/Notebook/13 July 2013

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13 July 2013

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Contents

208 lab


Main purpose today


Make gel purifications to enable USER-reactions and to make PCR of the fragments missing.

Who was in the lab


Kristian

Procedure


Purification gel 1,2 and 3

  • 1kb ladder
  • Nir
  • Nir
  • Nir
  • Nir
  • Nir
  • Nir
  • Nir
  • Nir
  • HAO

2013 07 13 gel Nir+HAO USER puri.jpg

  • 1kb ladder
  • cycAX
  • cycAX
  • cycAX
  • cycAX
  • AMO
  • AMO
  • AMO
  • AMO
  • HAO
  • HAO
  • HAO

J10893.jpg

  • 100 bp ladder, NEB
  • RFP
  • RFP
  • RFP

This is not a fucking duplicate.jpg

PCR of AMO

Did PCR on AMO on two programs: 1-3 on 54°C 4-6 on 52°C Both with the annealing time 3 min.

USER-reactions

There was made USER-reactions with all fragments after purification. AMO was purified even though it was only one very faint band; the others were bright band. The following plates where made:

  • RFP USER - RFP inside pZA21
  • AMO USER - AMO inside pZA21
  • HAO USER - HAO inside pZA21
  • cycAX USER - cycAX inside pZA21
  • Nir USER - Nir inside pZA21
  • Neg. USER - pZA21 linearized only

Conclusion


PCR of AMO with USER-primers was not a success we will see if the new PCR have done better. We will evaluate the transformants on Monday with colony-PCR.

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