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User:University of Washington/13 August 2008
From 2008.igem.org
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(→LuxR from AraC and TetR(Faifan)) |
(→LuxR from AraC and TetR(Faifan)) |
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<h4>Elowitz's</h4> | <h4>Elowitz's</h4> | ||
| + | |||
| + | -Made Glycerol Stock of BBa_Elowitz x 2 | ||
-Restriction Digest of BBa_Elowitz, BBa_E0240(GFP), BBa_I0462(LuxR), BBa_P1010(on pSB3K3) | -Restriction Digest of BBa_Elowitz, BBa_E0240(GFP), BBa_I0462(LuxR), BBa_P1010(on pSB3K3) | ||
| Line 43: | Line 45: | ||
*nanodrop at 230 nm | *nanodrop at 230 nm | ||
<table> | <table> | ||
| - | + | <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>76.2</td><td>1.12</td><td>0.34</td> | ||
| + | </tr> | ||
| + | <tr align="center"> | ||
| + | <td>LuxR</td><td>43.5</td><td>0.98</td><td>0.52</td> | ||
| + | </tr> | ||
| + | <tr align="center"> | ||
| + | <td>GFP</td><td>40.7</td><td>1.00</td><td>0.46</td> | ||
| + | </tr> | ||
| + | <tr align="center"> | ||
| + | <td>P1010</td><td>50.8</td><td>0.78</td><td>0.17</td> | ||
| + | </tr> | ||
</table> | </table> | ||
| - | + | *store in 4 degree Celsius | |
---- | ---- | ||
Back to [[Team:University_of_Washington/Notebook#Notebook]] | Back to [[Team:University_of_Washington/Notebook#Notebook]] | ||
Revision as of 22:22, 14 August 2008
Contents |
Conjugation (Scott)
experiment from Aug. 7 results
| experiment # | Bacteria | Yeast | Yeast count | transconjugation efficiency |
|---|---|---|---|---|
| 1 | 250uL | 250uL | 66 | 2.64E-5 |
| 2 | 200uL | 300uL | 64 | 2.13E-5 |
| 3 | 100uL | 400uL | 32 | 8.00e-6 |
| 4 | 50uL | 450uL | 18 | 4.00e-6 |
LuxR from AraC and TetR(Faifan)
QuikChange Approach
- Overnight QuikChange#4 culture from Amp plate (reaction1 x 4, reaction2 x 2) in Tsy+Amp
- Sequence of DNA from QuikChange#3 came back ==> No mutation.
Elowitz's
-Made Glycerol Stock of BBa_Elowitz x 2
-Restriction Digest of BBa_Elowitz, BBa_E0240(GFP), BBa_I0462(LuxR), BBa_P1010(on pSB3K3)
- mix
| dH2O(ul) | Buffer(ul) | BSA(ul) | DNA(ul) | E(ul) | X(ul) | S(ul) | P(ul) | |
|---|---|---|---|---|---|---|---|---|
| BBa_Elowitz | 32.5 | EcoRI, 5 | 0.5 | 10 | 1 | - | 1 | - |
| LuxR | 32.5 | NEB3, 5 | 0.5 | 10 | - | 1 | - | 1 |
| GFP | 32.5 | NEB3, 5 | 0.5 | 10 | - | 1 | - | 1 |
| P1010 | 32.5 | EcoRI, 5 | 0.5 | 10 | 1 | - | - | 1 |
- incubate 37 degree Celsius, 1.5 hours
- 15 mins water bath 80 degree Celsius
- nanodrop at 230 nm
| ng/ul | 260/280 | 260/230 | |
|---|---|---|---|
| BBa_Elowitz | 76.2 | 1.12 | 0.34 |
| LuxR | 43.5 | 0.98 | 0.52 |
| GFP | 40.7 | 1.00 | 0.46 |
| P1010 | 50.8 | 0.78 | 0.17 |
- store in 4 degree Celsius
Back to Team:University_of_Washington/Notebook#Notebook
