User:University of Washington/21 August 2008

From 2008.igem.org

(Difference between revisions)
 
(3 intermediate revisions not shown)
Line 1: Line 1:
 +
==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/ul260/280260/230
BBa_Elowitz2.11.560.03
LuxR5.81.980.06
GFP5.51.410.02
P1010(KAN)4.01.970.02

- ligation

  • mix
dH20(ul)Buffer(ul)P1010-Kan(ul)BBa_Elowitz(ul)GFP(ul)LuxR(ul)T4 ligase(ul)
GFP1.36253.427.22-1>/td>
LuxR1.2825.03.42-7.31>/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