Team:DTU-Denmark/Notebook/23 August 2013

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(Difference between revisions)
(PCR Biobrick Isolation)
(PCR constitutive reference promoter)
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===PCR constitutive reference promoter===
===PCR constitutive reference promoter===
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Got new primers for the constitutive reference promoter since the old reverse primer (52b2) had a mistake. Repeated reaction set-up and program from the 13.08. Used GC buffer, 5% DMSO and 2uL 50 mM MgCl2.
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primers: 52a, 52bn
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 +
template: pZA21::RFP
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program:
program:

Revision as of 15:47, 23 August 2013

23 August 2013

Contents

lab 208


Main purpose


Who was in the lab


Kristian, Henrike

Procedure


PCR Biobrick Isolation

Used new primers to isolate Hello World constructs. Used HF buffer and tested three different concentrations of DMSO: 0%, 2% and 5%.

Primers: 56a, 56b

template: TAT2-1 (miniprep)

program:

temperature time cycles
98C 2:00 -
98C 0:20 36
56C 0:30 36
72C 0:45 36
72C 5:00 -
10C hold -

PCR constitutive reference promoter

Got new primers for the constitutive reference promoter since the old reverse primer (52b2) had a mistake. Repeated reaction set-up and program from the 13.08. Used GC buffer, 5% DMSO and 2uL 50 mM MgCl2.

primers: 52a, 52bn

template: pZA21::RFP


program:

temperature time cycles
98C 2:00 -
98C 0:20 36
56C 1:00 36
72C 3:00 36
72C 5:00 -
10C hold -

Results


Gel electrophoresis

  • 1 kb ladder
  • Sec2 Biobrick
  • Sec2 Biobrick
  • Sec2 neg
  • col AMO 1
  • col AMO 2
  • col AMO 3
  • col AMO 4
  • col AMO 5
  • col AMO 6
  • col AMO 7
  • col AMO 8
  • col AMO 9
  • col AMO 10
  • 1 kb ladder

2013-08-23 bb sec amo colony.jpg

  • 1 kb ladder
  • col Nir
  • constitutive reference promoter
  • constitutive reference promoter
  • Biobrick Isolation TAT2-1 negative
  • Biobrick Isolation TAT2-1 2% DMSO
  • Biobrick Isolation TAT2-1 5% DMSO
  • Biobrick Isolation TAT2-1 w/o DMSO
  • 1 kb ladder

Conclusion


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