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User:University of Washington/9 September 2008
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| + | ==LuxR from AraC and TetR(Faifan)== | ||
| + | |||
| + | -Ran gel of the PCR products x14 colonies, got no DNA in all lanes-probably because cells weren't lysed or there weren't any cells at all from the beginning(resuspension wasn't good enough to disperse the cells in dH2O) | ||
| + | |||
| + | -Resuspended 28 colonies into 20 ul dH2O (14 from yesterday's resuspension, 14 from plate), boiled at 100 degree Celsius for 16 mins. | ||
| + | |||
| + | -Did Colony PCR again | ||
| + | *mix 12.6ul dH2O + 2 Buffer + 0.4ul 10mM-dNTP + 0.8ul 50mM-MgCl2 + 1ul 10mM-VF2 + 1ul 10mM-VR + 0.2ul Taq + 2ul Template | ||
| + | *Thermocycling | ||
| + | **1 : 95 : 1 min | ||
| + | **30: 95 : 30 s | ||
| + | **__: 53 : 30 s | ||
| + | **__: 72 : 1 min | ||
| + | **1 : 72 : 7 mins | ||
| + | **1 : 4 : forever | ||
| + | |||
| + | -Grew overnight of MG1655Z1 from stock in Tsy | ||
---- | ---- | ||
Back to [[Team:University_of_Washington/Notebook#Notebook]] | Back to [[Team:University_of_Washington/Notebook#Notebook]] | ||
Latest revision as of 22:57, 9 September 2008
LuxR from AraC and TetR(Faifan)
-Ran gel of the PCR products x14 colonies, got no DNA in all lanes-probably because cells weren't lysed or there weren't any cells at all from the beginning(resuspension wasn't good enough to disperse the cells in dH2O)
-Resuspended 28 colonies into 20 ul dH2O (14 from yesterday's resuspension, 14 from plate), boiled at 100 degree Celsius for 16 mins.
-Did Colony PCR again
- mix 12.6ul dH2O + 2 Buffer + 0.4ul 10mM-dNTP + 0.8ul 50mM-MgCl2 + 1ul 10mM-VF2 + 1ul 10mM-VR + 0.2ul Taq + 2ul Template
- Thermocycling
- 1 : 95 : 1 min
- 30: 95 : 30 s
- __: 53 : 30 s
- __: 72 : 1 min
- 1 : 72 : 7 mins
- 1 : 4 : forever
-Grew overnight of MG1655Z1 from stock in Tsy
Back to Team:University_of_Washington/Notebook#Notebook
