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User:University of Washington/7 August 2008
From 2008.igem.org
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- Inoculated overnight cultures with colony isolates of TOP10 for promoter constructs I20260, I20259, I20261, I20267, and empty TOP10 cells. | - Inoculated overnight cultures with colony isolates of TOP10 for promoter constructs I20260, I20259, I20261, I20267, and empty TOP10 cells. | ||
| + | |||
| + | ==LuxR from AraC and TetR(Faifan)== | ||
| + | |||
| + | - Both ligated DNA(Elowitz's promoter + pSB1AC3) transformation grew on Amp. Got single colonies. Picked and overnight. | ||
| + | |||
| + | - Continued QuikChange | ||
| + | *Added 1 ul dpn1 in reaction 1, 2 and negative control, mixed and span 1 min. | ||
| + | *Incubated 37 degree Celcius for 3 hours | ||
| + | *PCR purified | ||
| + | *Transformed 2 ul of each into XL1-Blue, grew on Amp plate | ||
| + | |||
| + | ==MG1655Z1== | ||
| + | |||
| + | -Diluted 1:100 of overnight culture of MG1655Z1(2 colonies from Tsy#2 plate) in Tsy broil. | ||
| + | |||
| + | -Incubated the dilution 3 hours | ||
| + | |||
| + | -Diluted 1:100 again in Tsy, inoculated in Tsy plates. | ||
| + | |||
---- | ---- | ||
Back to [[Team:University_of_Washington/Notebook#Notebook]] | Back to [[Team:University_of_Washington/Notebook#Notebook]] | ||
Revision as of 23:33, 7 August 2008
Contents |
Yeast Shuttle Plasmid
- Obtained yeast strain K. lactis
- Obtained primers for amplification of LAC4-LAC12 region of K. lactis genome
- PCR'd the K. lactis with primers
- Made glyercol stocks of BBa_J63005 and BBa_J33202
BioBrick Promoter Measurements
- Inoculated overnight cultures with colony isolates of TOP10 for promoter constructs I20260, I20259, I20261, I20267, and empty TOP10 cells.
LuxR from AraC and TetR(Faifan)
- Both ligated DNA(Elowitz's promoter + pSB1AC3) transformation grew on Amp. Got single colonies. Picked and overnight.
- Continued QuikChange
- Added 1 ul dpn1 in reaction 1, 2 and negative control, mixed and span 1 min.
- Incubated 37 degree Celcius for 3 hours
- PCR purified
- Transformed 2 ul of each into XL1-Blue, grew on Amp plate
MG1655Z1
-Diluted 1:100 of overnight culture of MG1655Z1(2 colonies from Tsy#2 plate) in Tsy broil.
-Incubated the dilution 3 hours
-Diluted 1:100 again in Tsy, inoculated in Tsy plates.
Back to Team:University_of_Washington/Notebook#Notebook
