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

From 2008.igem.org

(Difference between revisions)
(Thursday, 08/14/2008)
(Thursday, 08/14/2008)
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== Thursday, 08/14/2008 ==
== Thursday, 08/14/2008 ==
[[Image:HD 080814-LuxQ PCR.png|thumb|right|PCR for LuxQ and LuxQ-2]]
[[Image:HD 080814-LuxQ PCR.png|thumb|right|PCR for LuxQ and LuxQ-2]]
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* Gel from LuxQ, LuxQ
+
* Gel from LuxQ, LuxQ2
* sequentiell digestion of Fusion-1 with NcoI and NdeI (NEBuffer 3)
* sequentiell digestion of Fusion-1 with NcoI and NdeI (NEBuffer 3)
* Double digestion of pDK48 with NcoI/NdeI (NEBuffer 4)
* Double digestion of pDK48 with NcoI/NdeI (NEBuffer 4)

Revision as of 12:47, 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
Fusion-PCR for Fusion-1, LuxQ PCR, LuxQ-2 PCR, pDK48 miniprep and xbaI digestion of LuxQ no. 3
Digestion of Fusion-1 and pDK48 with NcoI/NdeI/AgeI


Thursday, 08/14/2008

PCR for LuxQ and LuxQ-2
  • Gel from LuxQ, LuxQ2
  • sequentiell digestion of Fusion-1 with NcoI and NdeI (NEBuffer 3)
  • Double digestion of pDK48 with NcoI/NdeI (NEBuffer 4)


Friday, 08/15/2008

Go to 3rd week