Team:DTU-Denmark/Notebook/2 August 2013

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(Difference between revisions)
(lab 208)
(lab 208)
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==Procedure==
==Procedure==
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===Gel purification===
* Gel purification was performed according to protocol included in QIAEX, Gel Extraction Kit.
* Gel purification was performed according to protocol included in QIAEX, Gel Extraction Kit.
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===USER reaction===
* USER reaction was performed according to standard protocol SOP1. We use DNA linear fragment containing pZA21 bacbone,cycAX with His-Tag sequence.
* USER reaction was performed according to standard protocol SOP1. We use DNA linear fragment containing pZA21 bacbone,cycAX with His-Tag sequence.
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===PCR ===
* PCR according to standard PCR protocol.  
* PCR according to standard PCR protocol.  
Samples names:  
Samples names:  
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* 9, 10 - Tat His-Tag, primers 20a, 20b; template: TAT 3 1a
* 9, 10 - Tat His-Tag, primers 20a, 20b; template: TAT 3 1a
* 11, 12, 13 - negative controls as follows for: Tat His-Tag, Nir, Sec His-Tag
* 11, 12, 13 - negative controls as follows for: Tat His-Tag, Nir, Sec His-Tag
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===Purification of plasmids after ethanol purification===
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According to protocol attached to the DNA purification kit, QIAGEN
==Results==
==Results==
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Revision as of 15:22, 2 August 2013

2 August 2013

Contents

lab 208


Main purpose


  • Gel purification of Cyc and Nir
  • USER reaction
  • PCR of Nir, His-Tag Sec, His-Tag TAT
  • Miniprep re-purification with columns

Who was in the lab


Gosia, Kristian, Natalia, Julia

Procedure


Gel purification

  • Gel purification was performed according to protocol included in QIAEX, Gel Extraction Kit.

USER reaction

  • USER reaction was performed according to standard protocol SOP1. We use DNA linear fragment containing pZA21 bacbone,cycAX with His-Tag sequence.

PCR

  • PCR according to standard PCR protocol.

Samples names:

  • 1,2 - Nir 48 (primers 48a, 48b) on template of Nir (too long fragment amplified at the beginning of work with Nir PCR)
  • 3,4 - Sec His-Tag, primers 19a, 19b; template: Sec 2
  • 5,6 - Tat His-Tag, primers 20a, 20b; template: TAT 2 1
  • 7,8 - Tat His-Tag, primers 20a, 20b; template: TAT 3 2
  • 9, 10 - Tat His-Tag, primers 20a, 20b; template: TAT 3 1a
  • 11, 12, 13 - negative controls as follows for: Tat His-Tag, Nir, Sec His-Tag

Purification of plasmids after ethanol purification

According to protocol attached to the DNA purification kit, QIAGEN

Results


Conclusion


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