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Team:Heidelberg/Notebook/Sensing Group/Notebook/6thweek
From 2008.igem.org
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Sequencing @ GATC yielded correct sequence for LuxQ no. 1 | Sequencing @ GATC yielded correct sequence for LuxQ no. 1 | ||
| - | * PCR for LuxQ and LuxP with Phusion Mastermix | + | * PCR for LuxQ and LuxP with Phusion (25 µl Mastermix, 0.5 µl each Primer, 0(1) µl DMSO, 24(23) µl H<sub>2</sub>0, 0.5 µl Template) |
| + | * PCR Programme: 5 min @ 98 °C || 10 s @ 98 °C | 30 s @ 55 °C | 30 s @ 72 °C || 5 min @ 72 °C | 4 °C | ||
[[Image:HD 080908-LuxQ P PCR.png|center|thumb|400px|PCR for LuxQ and LuxP. LuxQ PCR did not work, but first PCR for LuxP was positive.]] | [[Image:HD 080908-LuxQ P PCR.png|center|thumb|400px|PCR for LuxQ and LuxP. LuxQ PCR did not work, but first PCR for LuxP was positive.]] | ||
Revision as of 16:59, 26 October 2008
Contents |
Monday, 09/08/2008
- Liquid Culture of 5 colonies from LuxQ Transformation and 1 from F1 transformation
- Miniprep of Liquid Culture from previous Transformation (done by Chenchen). Incubated for two days @ RT and about 6 h @ 37 °C
- Digestion of the Miniprep products with xbaI (Buffer2 + BSA, 1 ul enzyme @ 37 °C about 40min)
Sequencing @ GATC yielded correct sequence for LuxQ no. 1
- PCR for LuxQ and LuxP with Phusion (25 µl Mastermix, 0.5 µl each Primer, 0(1) µl DMSO, 24(23) µl H20, 0.5 µl Template)
- PCR Programme: 5 min @ 98 °C || 10 s @ 98 °C | 30 s @ 55 °C | 30 s @ 72 °C || 5 min @ 72 °C | 4 °C
