Team:DTU-Denmark/Notebook/9 September 2013

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{{:Team:DTU-Denmark/Templates/StartPage|9 September}}
{{:Team:DTU-Denmark/Templates/StartPage|9 September}}
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=lab 208=
=lab 208=
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==Main purpose==
 
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==Who was in the lab==
==Who was in the lab==
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===Colony PCR to verify inserts HAO and Nir in pZA21::ara and pZA21::ara tight===
===Colony PCR to verify inserts HAO and Nir in pZA21::ara and pZA21::ara tight===
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Colony PCR on the Nir transformants, testing for NirH (primers 45a, 45b, annealing 59C), NirM+S (primers 44, 11a annealing 64C) and NirN (primers 43, 11b annealing 68C).
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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 || 10:00 || -
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|-
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|  98C || 0:10 || 36
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|-
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|  59C/64C/68C || 0:30 || 36
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|-
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|  72C || 2:00 || 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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|-
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|}
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Colony PCR to verify HAO insert.
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Primers: 28a, 28b
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temperature: 59C
==Results==
==Results==
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* colony 4, NirN  
* colony 4, NirN  
* 1 kb ladder
* 1 kb ladder
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[[File:2013-09-09 nir col.jpg|600px]]
The gel picture is inconclusive. We will do another colony PCR using the primers designed for sequencing.
The gel picture is inconclusive. We will do another colony PCR using the primers designed for sequencing.
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Navigate to the [[Team:DTU-Denmark/Notebook/8_September_2013|Previous]] or the [[Team:DTU-Denmark/Notebook/10_September_20133|Next]] Entry
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Navigate to the [[Team:DTU-Denmark/Notebook/8_September_2013|Previous]] or the [[Team:DTU-Denmark/Notebook/10_September_2013|Next]] Entry
{{:Team:DTU-Denmark/Templates/EndPage}}
{{:Team:DTU-Denmark/Templates/EndPage}}

Latest revision as of 12:13, 4 October 2013

9 September

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Contents

lab 208


Who was in the lab


Julia, Kristian, Henrike

Procedure


Colony PCR to verify inserts HAO and Nir in pZA21::ara and pZA21::ara tight

Colony PCR on the Nir transformants, testing for NirH (primers 45a, 45b, annealing 59C), NirM+S (primers 44, 11a annealing 64C) and NirN (primers 43, 11b annealing 68C).

program:

temperature time cycles
98C 10:00 -
98C 0:10 36
59C/64C/68C 0:30 36
72C 2:00 36
72C 5:00 -
10C hold -

Colony PCR to verify HAO insert.

Primers: 28a, 28b

temperature: 59C

Results


Transfomation

Transformation on last friday gave the following results:

  • HAO in pZA21::ara (from Biobrick K808000) - 6 colonies (3 orange and small, 3 normal color and very big)
  • negative HAO in pZA21::ara (from Biobrick K808000) - no colonies
  • HAO in pZA21::ara tight - a lot of colonies, some very big
  • negative HAO in pZA21::ara tight - a lot of small colonies
  • Nir in pZA21::ara tight - 8 colonies
  • negative Nir in pZA21::ara tight - no colonies


Gel

Gel on the results of the first colony PCR on the Nir transformants, testing for NirH, NirM+S and NirN

  • 1 kb ladder
  • colony 1, NirH
  • colony 1, Nir M+S
  • colony 1, NirN
  • colony 2, NirH
  • colony 2, Nir M+S
  • colony 2, NirN
  • colony 3, NirH
  • colony 3, Nir M+S
  • colony 3, NirN
  • colony 4, NirH
  • colony 4, Nir M+S
  • colony 4, NirN
  • 1 kb ladder

2013-09-09 nir col.jpg

The gel picture is inconclusive. We will do another colony PCR using the primers designed for sequencing.

Navigate to the Previous or the Next Entry