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User:University of Washington/21 August 2008
From 2008.igem.org
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==LuxR from AraC and TetR(Faifan)== | ==LuxR from AraC and TetR(Faifan)== | ||
| - | - | + | - nanodropped the gel purified DNA at 230 nm |
| + | <table border=1> | ||
| + | <tr align="center"> | ||
| + | <th></th><th>ng/ul</th><th>260/280</th><th>260/230</th> | ||
| + | </tr> | ||
| + | <tr align="center"> | ||
| + | <td>BBa_Elowitz</td><td>2.1</td><td>1.56</td><td>0.03</td> | ||
| + | </tr> | ||
| + | <tr align="center"> | ||
| + | <td>LuxR</td><td>5.8</td><td>1.98</td><td>0.06</td> | ||
| + | </tr> | ||
| + | <tr align="center"> | ||
| + | <td>GFP</td><td>5.5</td><td>1.41</td><td>0.02</td> | ||
| + | </tr> | ||
| + | <tr align="center"> | ||
| + | <td>P1010(KAN)</td><td>4.0</td><td>1.97</td><td>0.02</td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | |||
| + | - ligation | ||
| + | *mix | ||
| + | <table border=1> | ||
| + | <tr align="center"> | ||
| + | <th></th><th>dH20(ul)</th><th>Buffer(ul)</th><th>P1010-Kan(ul)</th><th>BBa_Elowitz(ul)</th><th>GFP(ul)</th><th>LuxR(ul)</th><th>T4 ligase(ul)</th> | ||
| + | </tr> | ||
| + | <tr align="center"> | ||
| + | <td>GFP</td><td>1.36</td><td>2</td><td>5</td><td>3.42</td><td>7.22</td><td>-</td><td>1>/td> | ||
| + | </tr> | ||
| + | <tr align="center"> | ||
| + | <td>LuxR</td><td>1.28</td><td>2</td><td>5.0</td><td>3.42</td><td>-</td><td>7.3</td><td>1>/td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | *1 hr incubate at room temp | ||
| + | *15 mins denature at 65 degree Celsius | ||
| + | *20 mins filtration | ||
| + | *transformed 2ul DNA into XL1-Blue, inoculated on Kan plate | ||
| + | |||
| + | ==MG1655Z1(Faifan)== | ||
| + | |||
| + | -None of the four cultures grew on Tet, Kan, Cam | ||
| + | |||
| + | -Only culture#3 didn't grow on Amp | ||
| + | *Streaked #3 to new Tsy plate | ||
| + | *grew overnight of #3 in Tsy | ||
| + | |||
| + | |||
| + | |||
| + | ==LuxR from pLac== | ||
| + | |||
| + | --I-insert and R-vector parts were isolated from their gel fragments and ligated together. | ||
| + | |||
---- | ---- | ||
Back to [[Team:University_of_Washington/Notebook#Notebook]] | Back to [[Team:University_of_Washington/Notebook#Notebook]] | ||
Latest revision as of 21:46, 22 August 2008
LuxR from AraC and TetR(Faifan)
- nanodropped the gel purified DNA at 230 nm
| ng/ul | 260/280 | 260/230 | |
|---|---|---|---|
| BBa_Elowitz | 2.1 | 1.56 | 0.03 |
| LuxR | 5.8 | 1.98 | 0.06 |
| GFP | 5.5 | 1.41 | 0.02 |
| P1010(KAN) | 4.0 | 1.97 | 0.02 |
- ligation
- mix
| dH20(ul) | Buffer(ul) | P1010-Kan(ul) | BBa_Elowitz(ul) | GFP(ul) | LuxR(ul) | T4 ligase(ul) | |
|---|---|---|---|---|---|---|---|
| GFP | 1.36 | 2 | 5 | 3.42 | 7.22 | - | 1>/td> |
| LuxR | 1.28 | 2 | 5.0 | 3.42 | - | 7.3 | 1>/td> |
- 1 hr incubate at room temp
- 15 mins denature at 65 degree Celsius
- 20 mins filtration
- transformed 2ul DNA into XL1-Blue, inoculated on Kan plate
MG1655Z1(Faifan)
-None of the four cultures grew on Tet, Kan, Cam
-Only culture#3 didn't grow on Amp
- Streaked #3 to new Tsy plate
- grew overnight of #3 in Tsy
LuxR from pLac
--I-insert and R-vector parts were isolated from their gel fragments and ligated together.
Back to Team:University_of_Washington/Notebook#Notebook
