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Team:Heidelberg/Notebook/Sensing Group/Notebook/3rdweek
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* Fusion-PCR for Fusion-1 and Fusion-2, each 4 tubes (2 µl Tar mixed with 1 µl LuxQ) | * Fusion-PCR for Fusion-1 and Fusion-2, each 4 tubes (2 µl Tar mixed with 1 µl LuxQ) | ||
* preparation of O/N culture of LuxS and pDK48 transformed cells for subsequent miniprep | * preparation of O/N culture of LuxS and pDK48 transformed cells for subsequent miniprep | ||
| - | * digestion of pDK48 with NcoI/NdeI | + | * digestion of pDK48 with NcoI/NdeI (NEBuffer 4) |
| - | * digestion of pTrc99alpha and LuxQ | + | * digestion of pTrc99alpha and LuxQ with BamHI/NcoI (NEBuffer 3 + BSA) and gel extraction |
== Tuesday, 08/19/2008 == | == Tuesday, 08/19/2008 == | ||
Revision as of 23:35, 26 October 2008
Monday, 08/18/2008
- PCR again for LuxQ-1 and LuxQ-2 for Fusion constructs. Different Mg concentrations from 1% to 4% were used. For LuxQ-2 another PCR was run using as template V. harveyi colonies
- Fusion-PCR for Fusion-1 and Fusion-2, each 4 tubes (2 µl Tar mixed with 1 µl LuxQ)
- preparation of O/N culture of LuxS and pDK48 transformed cells for subsequent miniprep
- digestion of pDK48 with NcoI/NdeI (NEBuffer 4)
- digestion of pTrc99alpha and LuxQ with BamHI/NcoI (NEBuffer 3 + BSA) and gel extraction
Tuesday, 08/19/2008
- Miniprep of pDK48 and LuxS
- gel of Fusion constructs and gelextraction of pDK48 from yesterday
- afterwards another gel was run to purify (get rid of the primers used) the Fusion constructs via gel extraction
Wednesday, 08/20/2008 - Friday, 08/22/2008
- nothing important to report
