Team:Heidelberg/Notebook/Sensing Group/Notebook/2ndweek

From 2008.igem.org

(Difference between revisions)
(Wednesday, 08/13/2008)
(Wednesday, 08/13/2008)
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* Miniprep of LuxQ no. 3 O/N culture and digestion with xbaI
* Miniprep of LuxQ no. 3 O/N culture and digestion with xbaI
* Fusion-PCR for Fusion-1 and repetetion of PCR for LuxQ-2 and LuxQ
* Fusion-PCR for Fusion-1 and repetetion of PCR for LuxQ-2 and LuxQ
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[[Image:HD 080813-LuxQ PCR-pDK48 mini LuxQ3 xbaI.png|center|thumb|500px|Fusion-PCR for Fusion-1, LuxQ PCR, LuxQ-2 PCR, pDK48 miniprep and xbaI digestion of LuxQ no. 3]]
 
* digestion of pDK48 and Fusion-1 with NcoI/NdeI (0.5 µl) and AgeI (1 µl) in NEBuffer 4
* digestion of pDK48 and Fusion-1 with NcoI/NdeI (0.5 µl) and AgeI (1 µl) in NEBuffer 4
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[[Image:HD 080813-F1 pDK48 Digestion.png|center|thumb|200px|Digestion of Fusion-1 and pDK48 with NcoI/NdeI/AgeI]]
+
[[Image:HD 080813-F1 pDK48 Digestion.png|right|thumb|200px|Digestion of Fusion-1 and pDK48 with NcoI/NdeI/AgeI]]
 +
[[Image:HD 080813-LuxQ PCR-pDK48 mini LuxQ3 xbaI.png|thumb|500px|Fusion-PCR for Fusion-1, LuxQ PCR, LuxQ-2 PCR, pDK48 miniprep and xbaI digestion of LuxQ no. 3]]
== Thursday, 08/14/2008 ==
== Thursday, 08/14/2008 ==

Revision as of 10:55, 26 October 2008

Back to the overview

Contents

Monday, 08/11/2008

  • preparation of LuxS harboring cells O/N culture

Tuesday, 08/12/2008

  • Miniprep of correctly sequenced LuxS and Glycerol-stock
  • LuxQ no. 3 checked via digestion with NcoI/BamHI --> negative result
  • PCR for LuxQ, LuxQ-1, LuxQ-2, Tar-1, Tar-2 with Taq Mastermix
  • preparation of O/N culture for LuxQ no. 3

Wednesday, 08/13/2008

  • PCR-purification of the 5 PCR-products eluted in 50 µl H2O
  • Miniprep of LuxQ no. 3 O/N culture and digestion with xbaI
  • Fusion-PCR for Fusion-1 and repetetion of PCR for LuxQ-2 and LuxQ
  • digestion of pDK48 and Fusion-1 with NcoI/NdeI (0.5 µl) and AgeI (1 µl) in NEBuffer 4
Digestion of Fusion-1 and pDK48 with NcoI/NdeI/AgeI
Fusion-PCR for Fusion-1, LuxQ PCR, LuxQ-2 PCR, pDK48 miniprep and xbaI digestion of LuxQ no. 3

Thursday, 08/14/2008

Friday, 08/15/2008

Go to 3rd week