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User:University of Washington/6 August 2008
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| + | ==LuxR from AraC and TetR(Faifan)== | ||
| + | |||
| + | -Tranformation of ligation product at room temp(P1010 on pSB1AC3 and promoter) succeed? There were too much growth, so the culture was streaked out in new Amp plate. | ||
| + | |||
| + | -Continued ligation process (the one at 4 degree Celcius overnight): denatured enzyme, filterd, tranformed into XL1-Blue, and grew on Amp plate. | ||
| + | |||
| + | -Discovered today that the dsDNA added for previous QuikChange reaction was way excessive due to unit misunderstanding. | ||
| + | |||
| + | -Did first part of QuikChange (set up reaction + theymocycling overnight) | ||
| + | *nanodropped R0080(AraC) = 113.5 ng/ul, 1.9-260/280, 2.19-260/230 | ||
| + | *Elongation step 68 degree Celcius for 7 mins was removed in theymocycling. | ||
| + | |||
| + | ==MG1655Z1(Faifan)== | ||
| + | |||
| + | -Got growth in Amp plate from MG1655Z1 stock. Have to find out what plasmid contains Amp resistance from literature. | ||
| + | |||
| + | -Got growth in Amp and a bit in Tet plates from MG1655Z1 Tsy#2 plate. This said that there were still some unknown plasmids in the strain. Two colonies from section 3 and 5 were picked and grew overnight in Tsy. | ||
| + | |||
| + | |||
---- | ---- | ||
Back to [[Team:University_of_Washington/Notebook#Notebook]] | Back to [[Team:University_of_Washington/Notebook#Notebook]] | ||
Revision as of 00:44, 7 August 2008
LuxR from AraC and TetR(Faifan)
-Tranformation of ligation product at room temp(P1010 on pSB1AC3 and promoter) succeed? There were too much growth, so the culture was streaked out in new Amp plate.
-Continued ligation process (the one at 4 degree Celcius overnight): denatured enzyme, filterd, tranformed into XL1-Blue, and grew on Amp plate.
-Discovered today that the dsDNA added for previous QuikChange reaction was way excessive due to unit misunderstanding.
-Did first part of QuikChange (set up reaction + theymocycling overnight)
- nanodropped R0080(AraC) = 113.5 ng/ul, 1.9-260/280, 2.19-260/230
- Elongation step 68 degree Celcius for 7 mins was removed in theymocycling.
MG1655Z1(Faifan)
-Got growth in Amp plate from MG1655Z1 stock. Have to find out what plasmid contains Amp resistance from literature.
-Got growth in Amp and a bit in Tet plates from MG1655Z1 Tsy#2 plate. This said that there were still some unknown plasmids in the strain. Two colonies from section 3 and 5 were picked and grew overnight in Tsy.
Back to Team:University_of_Washington/Notebook#Notebook
