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User:University of Washington/26 August 2008
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| + | ==LuxR from AraC and TetR(Faifan)== | ||
| + | |||
| + | <h4>ligation from gel extraction</h4> | ||
| + | -cultures of plasmid construct transformation ended up growing on Kan plates after incubating overnight at 36 degree Celsius | ||
| + | |||
| + | -selected 3 colonies from each plate(LuxR, GFP) and grew overnight to do sequencing | ||
| + | |||
| + | <h4>new ligation with CIP reaction</h4> | ||
| + | -miniprep I0462 x 3, ran gel and combined tubes | ||
| + | |||
| + | -restriction digest GFP and LuxR as in the table from yesterday(8/25) | ||
| + | |||
| + | -PCR purified digested promoter | ||
| + | |||
| + | -nanodropped the purified promoter: 261.9 ng/ul, 1.87(260/280), 2.04(260/230) | ||
| + | |||
| + | -did CIP reaction (Ingrid) | ||
| + | *mix 10X CIP Buffer + 0.5 units of CIP per ul/DNA + DNA(have final concentration of 0.5 ul/10 ul) | ||
| + | *incubated 37 degree Celsius 1 hour | ||
| + | *stored at -20 | ||
| + | |||
| + | ==MG1655Z1(Faifan)== | ||
| + | |||
| + | -made glycerol stock of the strain. (non-resistance) | ||
| + | |||
---- | ---- | ||
Back to [[Team:University_of_Washington/Notebook#Notebook]] | Back to [[Team:University_of_Washington/Notebook#Notebook]] | ||
Latest revision as of 20:23, 26 August 2008
Contents |
LuxR from AraC and TetR(Faifan)
ligation from gel extraction
-cultures of plasmid construct transformation ended up growing on Kan plates after incubating overnight at 36 degree Celsius
-selected 3 colonies from each plate(LuxR, GFP) and grew overnight to do sequencing
new ligation with CIP reaction
-miniprep I0462 x 3, ran gel and combined tubes
-restriction digest GFP and LuxR as in the table from yesterday(8/25)
-PCR purified digested promoter
-nanodropped the purified promoter: 261.9 ng/ul, 1.87(260/280), 2.04(260/230)
-did CIP reaction (Ingrid)
- mix 10X CIP Buffer + 0.5 units of CIP per ul/DNA + DNA(have final concentration of 0.5 ul/10 ul)
- incubated 37 degree Celsius 1 hour
- stored at -20
MG1655Z1(Faifan)
-made glycerol stock of the strain. (non-resistance)
Back to Team:University_of_Washington/Notebook#Notebook
