Team:DTU-Denmark/Notebook/29 July 2013

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(Procedure)
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PCR programms based on [[Team:DTU-Denmark/Methods/PCR| standard PCR programm]] with annealing temperatures 52C for HAO and 56C for AMO and extension time 3:00 mins.
PCR programms based on [[Team:DTU-Denmark/Methods/PCR| standard PCR programm]] with annealing temperatures 52C for HAO and 56C for AMO and extension time 3:00 mins.
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===Colony PCR for Nir===
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In order to extract Nir from ''Pseudomonas'' we used primers which do not contain uracil and new PCR programm (touch-down PCR). Phusion polymerase was used.
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Names of samples are as follows:
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1,2 - primers 41a, 41b - Nir, part 1
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3,4 - primers 42a, 42b - Nir, part 2
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5,6 - primers 11a, 11b - all fragment
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PCR programm (write it later when programm will be done)
==Results==
==Results==

Revision as of 12:14, 29 July 2013

29 July 2013

Contents

lab 208


Main purpose


  • Colony PCR from Pseudomonas aeruginosa (PAO) to isolate Nir.
  • Colony PCR to check the presence of AMO and HAO in transormants E.coli.

Who was in the lab


Kristian, Gosia, Henrike, Julia

Procedure


Colony PCR

Colony PCR was performed according to standard method in order to check if transformed cells which grew on medium contain desired insert in pZA21 plasmid. The colonies were chosen from agar plate to be checked. Names and numbers of colonies as well as plates names and primers are:

  • HAO 1-6 in pZA21 USER, primers 28a, 28b
  • AMO 1-9 in pZA21 USER, primers 29a, 29b


PCR programms based on standard PCR programm with annealing temperatures 52C for HAO and 56C for AMO and extension time 3:00 mins.

Colony PCR for Nir

In order to extract Nir from Pseudomonas we used primers which do not contain uracil and new PCR programm (touch-down PCR). Phusion polymerase was used.

Names of samples are as follows: 1,2 - primers 41a, 41b - Nir, part 1 3,4 - primers 42a, 42b - Nir, part 2 5,6 - primers 11a, 11b - all fragment

PCR programm (write it later when programm will be done)

Results


Conclusion


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