Team:DTU-Denmark/Notebook/16 August 2013

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(→Gradient PCR for Nir2 USER)
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*templates: TAT3-2, Sec2, cycAX
*templates: TAT3-2, Sec2, cycAX
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Used two different temperatures. Program:
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Tried two different annealing temperatures. Program:
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{| class="wikitable" style="text-align: right"
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! temperature !! time !! cycles
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|-
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|  98C || 2:00 || -
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|-
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|  98C || 0:10 || 36
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|-
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|  60C/56C || 0:45 || 36
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|-
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|  72C || 0:45 || 36
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|-
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|  72C || 5:00 || -
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|-
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|  10C || hold || -
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|-
 +
|}
==Results==
==Results==

Revision as of 12:01, 16 August 2013

16 August 2013

Contents

lab 208

Main purpose

Who was in the lab

Kristian, Julia, Henrike

Procedure

Gradient PCR for Nir2 USER

Set up new gradient PCR for Nir2 USER to run in 223. Reaction mix without MgCl2 but using DMSO and GC-buffer.

  • template: Nir2 extraction fragment
  • primers: 40a, 40b
  • additives: 5% DMSO
  • buffer: GC

Gradient went from 60C to 72C in 12 steps.

sample # 1 2 3 4 5 6 7 8 9 10 11 12
temperature 60.2 60.5 61.6 63.0 64.2 65.4 66.6 67.6 68.9 70.2 71.6 72

PCR for BioBrick construction

PCR amplified the constructs from Hello World and cytochromes and pSB1C3, which will be ligated together into biobricks.

constructs:

  • GC-buffer, 5%DMSO, 2uL 50mM MgCl2
  • primers: 53a, 53b
  • templates: TAT3-2, Sec2, cycAX

Tried two different annealing temperatures. Program:

temperature time cycles
98C 2:00 -
98C 0:10 36
60C/56C 0:45 36
72C 0:45 36
72C 5:00 -
10C hold -

Results

Conclusion

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