Team:DTU-Denmark/Notebook/10 July 2013

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(→Main purpose)
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* step 6: 90 sec (instead of 60 s)at 42 <sup>o</sup>C  
* step 6: 90 sec (instead of 60 s)at 42 <sup>o</sup>C  
* step 9: Incubation without shaking
* step 9: Incubation without shaking
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===Extraction PCR===
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9 samples from culture pAO1
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10 reactions
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* dNTP: 1 ul (10 ul)
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* HF buffer: 10 ul (100 ul)
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* Phusion polymerase: 0.5 ul (5 ul)
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* H<sub>2</sub>O: 31.5 ul (315 ul)
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Settings for PCR
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{| class="wikitable sortable" style="text-align: right"
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! Temperature (<sup>o</sup>C)!! Time (min)!! Rounds
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|-
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| 98 || 2:00 || 1
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|-
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| 98 || 0:20 || 35
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|-
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| 66 || 0:45 || 35
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|-
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| 72 || 2:00 || 35
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|-
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| 72 || 5:00 || 1
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|-
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| 4|| ∞ || -
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|}
==Results==
==Results==

Revision as of 16:21, 10 July 2013

Contents

208

Main purpose

Run gel with 9 PCR samples of Nir operon from Pseudomonas aeruginosa

Transformation of Biobricks

PCR reaction for Nir operon

Who was in the lab

Ariadni,Henrike,Julia,Natalia

Procedure

Run gel

  • 6 samples from PCR of 09.07.2013
  • ladder broadband
  • 3 samples from purification of 09.07.2013

Transformation of Biobricks

According to the iGEM protocol (transformation protocol) with slight changes.

  • step 1: 100 μl cells
  • step 2: 1.5 μl of the resuspended DNA
  • step 6: 90 sec (instead of 60 s)at 42 oC
  • step 9: Incubation without shaking

Extraction PCR

9 samples from culture pAO1 10 reactions

  • dNTP: 1 ul (10 ul)
  • HF buffer: 10 ul (100 ul)
  • Phusion polymerase: 0.5 ul (5 ul)
  • H2O: 31.5 ul (315 ul)

Settings for PCR

Temperature (oC) Time (min) Rounds
98 2:00 1
98 0:20 35
66 0:45 35
72 2:00 35
72 5:00 1
4 ∞ -

Results

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