Team:Osaka/NOTES

From 2009.igem.org

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     <h2>Registry Parts</h2></div>  
     <h2>Registry Parts</h2></div>  
   <div class="CollapsiblePanelContent">  
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<h3>Osaka team ALL PARTS  <a href="https://2009.igem.org/Team:Osaka/Parts_%26_Devices">here</a>Parts & Devices</h3>
+
<h3>All of Osaka's Parts and Devices is <a href="https://2009.igem.org/Team:Osaka/Parts_%26_Devices">here</a></h3>
<h4>COLOR</h4>
<h4>COLOR</h4>
<br>
<br>
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     <h2>Lab Note</h2></div>  
     <h2>Lab Note</h2></div>  
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 +
  <h3>Wet Lab</h3>
-
  <h4><p>Wet Lab</p></h4>
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<p>Week 1 : 8/2~8/8</p>
-
 
+
<p>Brainstorming , parts planning , prepared competent cell</p>
-
Week 1 : 8/2~8/8<br>
+
<p>ransformation and miniprep from MIT plate</p>
-
Brainstorming , parts planning , prepared competent cell<br>
+
-
transformation and miniprep from MIT plate<br>
+
-
 
+
<br>
<br>
-
week 2 : 8/9~8/15<br>
+
<p>week 2 : 8/9~8/15</p>
-
start assembly parts (K204001 ~ K204006) , transformation and mini prep from MIT
+
<p>start assembly parts (K204001 ~ K204006) , transformation and mini prep from MIT</p>
<br>
<br>
-
 
+
<p>Week 3 : 8/16~8/22</p>
 +
<p>assembly parts (~K204015)  ,  parts planning</p>
<br>
<br>
-
Week 3 : 8/16~8/22<br>
+
<p>Week 4 : 8/23~8/29</p>
-
assembly parts (~K204015) ,  parts planning<br>
+
<p>assembly parts (~K204021)</p>
-
 
+
<br>
<br>
-
Week 4 : 8/23~8/29<br>
+
<p>Week 5 : 8/30~9/5</p>
-
assembly parts (~K204021)<br>
+
<p>assembly parts (~K204030)</p>
-
 
+
<br>
<br>
-
Week 5 : 8/30~9/5<br>
+
<p>Week 6 : 9/6~9/12</p>
-
assembly parts (~K204030)<br>
+
<p>assembly parts (~K204046)</p>
-
 
+
<br>
<br>
-
Week 6 : 9/6~9/12<br>
+
<p>Week 7 : 9/13~9/19</p>
-
assembly parts (~K204046)<br>
+
<p>assembly parts (~K204050)</p>
-
 
+
<br>
<br>
-
Week 7 : 9/13~9/19<br>
+
<p>Week 8 : 9/20~9/26</p>
-
assembly parts (~K204050)<br>
+
<p>assembly parts (~K204056)</p>
-
 
+
<p>TEST : sensitivity of inducer and receptor</p>
 +
<p>Sample : 3O<sub>6</sub>HSL , C<sub>4</sub>HSL , AI-1 , 3OH,C<sub>14</sub>:1-HSL</p>
 +
<p>Result : Not confirm the fluorescence...</p>
<br>
<br>
-
Week 8 : 9/20~9/26<br>
+
<p>Week 9 : 9/27~10/3</p>
-
assembly parts (~K204056)<br>
+
<p>assembly parts (~K204066 , X1~X8), color (fluorescence) intensity check</p>
-
TEST : sensitivity of inducer and receptor <br>
+
-
Sample : 3O<sub>6</sub>HSL , C<sub>4</sub>HSL , AI-1 , 3OH,C<sub>14</sub>:1-HSL<br>
+
-
Result : Not confirm the fluorescence...<br>
+
-
 
+
-
 
+
<br>
<br>
-
Week 9 : 9/27~10/3<br>
+
<p>Week 10 : 10/4~10/10</p>
-
assembly parts (~K204066 , X1~X8) , color (fluorescence) intensity check<br>
+
<p>assembly parts (~K204078)</p>
-
 
+
<p>Lux receiver sensitivity test, color (fluorescence) intensity check</p>
-
 
+
<p>sequence check : generator and receiver</p>
 +
<p>3O<sub>6</sub>HSL , C<sub>4</sub>HSL , AI-1 , 3OH,C<sub>14</sub>:1-HSL</p>
 +
<h4 style="text-indent:2em">Get the parts from Team Chiba. Thanks a lot!!</h4>
<br>
<br>
-
Week 10 : 10/4~10/10<br>
+
<p>Week 11 : 10/11~10/17</p>
-
assembly parts (~K204078)<br>
+
<p>assembly parts (~K2040104)</p>
-
Lux receiver sensitivity test  ,  color (fluorescence) intensity check<br>
+
<p>sequence check : FliH etc…</p>
-
sequence check : generator and receiver<br>
+
<p>send Osaka team parts to MIT</p>
-
3O<sub>6</sub>HSL , C<sub>4</sub>HSL , AI-1 , 3OH,C<sub>14</sub>:1-HSL<br>
+
<p>FINISH!</p>
-
 
+
<h4 style="text-indent:2em">Finally we assembled about 120 parts!!</h4>
-
Get the parts from the Chiba University Team. , Thanks chiba team
+
<br>
<br>
-
 
+
<h3>Dry Lab</h3>
 +
<p>Week 5 : 8/30~9/5</p>
 +
<p>Planing model.</p>
<br>
<br>
-
Week 11 : 10/11~10/17<br>
+
<p>Week 6 : 9/6~9/12</p>
-
assembly parts (~K2040104)<br>
+
<p>Studying programing using Matlab. </p>
-
sequence check : FliH etc…<br>
+
-
send Osaka team parts to MIT<br>
+
-
FINISH!<br>
+
-
<b>Osaka team assembled about 120 parts!</b><br>
+
-
 
+
<br>
<br>
-
<h4><p>Dry Lab</p></h4>
+
<p>Week 7 : 9/13~9/19</p>
-
Week 5 : 8/30~9/5<br>
+
<p>Making program in case of single coloney.</p>
-
Planing model.<br>
+
-
 
+
<br>
<br>
-
Week 6 : 9/6~9/12<br>
+
<p>Week 8 : 9/20~9/26</p>
-
Studying programing using Matlab. <br>
+
<p>Making program in case of more than two colonies</p>
-
 
+
<p>Making movie file</p>
 +
<p>Searching paper written some parameter</p>
<br>
<br>
-
Week 7 : 9/13~9/19<br>
+
<p>Week 9 : 9/27~10/3</p>
-
Making program in case of single coloney.<br>
+
<p>Blash up our program </p>
-
 
+
<p>Searching paper written some parameter</p>
<br>
<br>
-
Week 8 : 9/20~9/26<br>
+
<p>Week 10 : 10/4~10/10</p>
-
Making program in case of more than two colonies<br>
+
<p>Making program of color gradation</p>
-
Making movie file<br>
+
-
Searching paper written some parameter<br>
+
-
 
+
<br>
<br>
-
Week 9 : 9/27~10/3<br>
+
<p>Week 11 : 10/11~10/17</p>
-
Blash up our program <br>
+
<p>Blash up our program</p>
-
Searching paper written some parameter<br>
+
-
 
+
<br>
<br>
-
Week 10 : 10/4~10/10<br>
+
<h3>Atelie</h3>
-
Making program of color gradation<br>
+
<p>Week 8 : 9/20~9/26</p>
-
 
+
<p>Making art works</p>
<br>
<br>
-
Week 11 : 10/11~10/17<br>
+
<p>Week 9 : 9/27~10/3</p>
-
Blash up our program<br>
+
<p>Making art works</p>
<br>
<br>
-
<h4><p>Atelie</p></h4>
+
<p>Week 10 : 10/4~10/10</p>
-
Week 8 : 9/20~9/26<br>
+
<p>Making art works</p>
-
Making art works<br>
+
-
 
+
<br>
<br>
-
Week 9 : 9/27~10/3<br>
+
<p>Week 11 : 10/11~10/17</p>
-
Making art works<br>
+
<p>Making art works</p>
-
 
+
-
<br>
+
-
Week 10 : 10/4~10/10<br>
+
-
Making art works<br>
+
-
 
+
-
<br>
+
-
Week 11 : 10/11~10/17<br>
+
-
Making art works<br>
+
-
 
+
-
 
+
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   </div>  
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    <h2>Protocols</h2></div>
 
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  <div class="CollapsiblePanelContent">
 
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    <p>Sensor Experiment 1: Receivers vs AHL<br>
 
-
 
-
Day 1 - Preparation<br>
 
-
1. From transformation plate, pick up a small quantity of cells from a colony using sterilized toothpick and streak on LA plate with the appropriate antibiotic resistance. Incubate at 37 degrees Celsius for 8+ hr.<br>
 
-
 
-
2. From the streak plate, choose a single-cell colony and pick up cells using sterilized toothpick, then inoculate 5ml LB culture medium to which the appropriate antibiotic resistance (50mg/l) has been added. Incubate overnight at 37 degrees Celsius in a shaking incubator.<br>
 
-
 
-
 
-
<br>
 
-
Day 2 - Measurement<br>
 
-
1. Transfer 1ml of the overnight culture into 30ml fresh LB culture medium. Incubate in shaking incubator at 37 degrees.<br>
 
-
<br>
 
-
2. Measure OD of culture every hour or so, until OD reaches fixed value (culture at full growth).<br>
 
-
<br>
 
-
3. Transfer culture into 50ml ultracentrifuge tube and spin at 8000rpm for 5 minutes.<br>
 
-
<br>
 
-
4. Resuspend pellet with 30ml fresh LB culture.<br>
 
-
<br>
 
-
5. Incubate culture in shaking incubator at 30 degree Celcius for 10 minutes.<br>
 
-
<br>
 
-
6. Transfer 1ml of culture into microcentrifuge tube and put on ice. Immediately add AHL to rest of culture to attain desired concentration.<br>
 
-
<br>
 
-
(Note: Between the steps detailed below, samples should be kept on ice to minimize cellular activity.)<br>
 
-
<br>
 
-
7. Obtain OD600 of 2x diluted sample by transferring 500ul of sample into cuvette, adding 500ul MiliQ water and measuring the resulting OD.<br>
 
-
<br>
 
-
8. Ultracentrifuge remaining 500ul of sample at 13000rpm for 2min, discard supernatant, then resuspend pellet with 1ml MiliQ water.<br>
 
-
<br>
 
-
9. Ultracentrifuge sample again at 13000rpm for 2min, discard supernatant and resuspend with precisely 1ml MiliQ water (this results in 2x dilution).<br>
 
-
<br>
 
-
10. Measure GFP fluorescence with a fluorimeter at 488nm excitation, 513nm emission.<br>
 
-
<br>
 
-
11. Every hour, sample 1ml of culture and repeat steps 7-10 until fluorescence reaches steady state.<br>
 
-
<br>
 
-
<br>
 
-
 
-
Sensor Experiment 2: Senders vs Receivers<br>
 
-
 
-
Day 1 - Preparation<br>
 
-
1. From transformation plate, pick up a small quantity of cells from a colony using sterilized toothpick and streak on LA plate with the appropriate antibiotic resistance. Incubate at 37 degrees Celsius for 8+ hr.<br>
 
-
<br>
 
-
2. From the streak plate, choose a single-cell colony and pick up cells using sterilized toothpick, then inoculate 5ml LB culture medium to which the appropriate antibiotic resistance (50mg/l) has been added. Incubate overnight at 37 degrees Celsius in a shaking incubator.<br>
 
-
<br>
 
-
 
-
Day 2 - Mix & Culture<br>
 
-
1. Prepare microcentrifuge tubes: 3 samples for each sender-receiver pair (sender-receiver test) or AHL concentration (receiver-only test). Add 400ul of LB-Amp culture medium to each microcentrifuge tube.<br>
 
-
<br>
 
-
2. Add AHL to each microcentrifuge tube that will be used for receiver test.<br>
 
-
<br>
 
-
3. Transfer 1ml of each overnight culture to separate microcentrifuge tubes and ultracentrifuge at 12000rpm for 1min. Discard supernatant and resuspend cells with 1ml LB-Amp. Repeat with remaining overnight culture if necessary to obtain the required amount of cell culture.<br>
 
-
<br>
 
-
4. For sender-receiver test: Add 50ul each of sender and receiver cultures to a microcentrifuge tube prepared in step 1. For receiver-only test: Add 100ul of receiver culture to a microcentrifuge tube that has been prepared in steps 1 & 2 (AHL added).<br>
 
-
<br>
 
-
5. Incubate microcentrifuge tubes at 37 degrees Celsius overnight.<br>
 
-
<br>
 
-
 
-
Day 3 - Observation & Measurement<br>
 
-
1. Wash cells by ultracentrifuging at 13000rpm for 2min, discarding supernatant and resuspending cells with MiliQ water. Repeat, and resuspend with precisely 1ml MiliQ water.<br>
 
-
<br>
 
-
2. For direct observation, bring cells suspended in water to darkened room and shine with UV Black Light. GFP fluorescence should be observable if sufficiently strong promoter activation has been achieved.<br>
 
-
<br>
 
-
3. Measure GFP fluorescence with a fluorimeter at 488nm excitation, 513nm emission.<br>
 
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<br>
 
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Latest revision as of 18:39, 21 October 2009

Home of iGEMOSAKA wiki

NOTES

Calendar

July
MTWTFSS
    [http://2009.igem.org/wiki/index.php?title=July/1_July_2009&action=edit 1] [http://2009.igem.org/wiki/index.php?title=July/2_July_2009&action=edit 2] [http://2009.igem.org/wiki/index.php?title=July/3_July_2009&action=edit 3] [http://2009.igem.org/wiki/index.php?title=July/4_July_2009&action=edit 4] [http://2009.igem.org/wiki/index.php?title=July/5_July_2009&action=edit 5]
[http://2009.igem.org/wiki/index.php?title=July/6_July_2009&action=edit 6] [http://2009.igem.org/wiki/index.php?title=July/7_July_2009&action=edit 7] [http://2009.igem.org/wiki/index.php?title=July/8_July_2009&action=edit 8] [http://2009.igem.org/wiki/index.php?title=July/9_July_2009&action=edit 9] [http://2009.igem.org/wiki/index.php?title=July/10_July_2009&action=edit 10] [http://2009.igem.org/wiki/index.php?title=July/11_July_2009&action=edit 11] [http://2009.igem.org/wiki/index.php?title=July/12_July_2009&action=edit 12]
[http://2009.igem.org/wiki/index.php?title=July/13_July_2009&action=edit 13] [http://2009.igem.org/wiki/index.php?title=July/14_July_2009&action=edit 14] [http://2009.igem.org/wiki/index.php?title=July/15_July_2009&action=edit 15] [http://2009.igem.org/wiki/index.php?title=July/16_July_2009&action=edit 16] [http://2009.igem.org/wiki/index.php?title=July/17_July_2009&action=edit 17] [http://2009.igem.org/wiki/index.php?title=July/18_July_2009&action=edit 18] [http://2009.igem.org/wiki/index.php?title=July/19_July_2009&action=edit 19]
[http://2009.igem.org/wiki/index.php?title=July/20_July_2009&action=edit 20] [http://2009.igem.org/wiki/index.php?title=July/21_July_2009&action=edit 21] [http://2009.igem.org/wiki/index.php?title=July/22_July_2009&action=edit 22] [http://2009.igem.org/wiki/index.php?title=July/23_July_2009&action=edit 23] [http://2009.igem.org/wiki/index.php?title=July/24_July_2009&action=edit 24] [http://2009.igem.org/wiki/index.php?title=July/25_July_2009&action=edit 25] [http://2009.igem.org/wiki/index.php?title=July/26_July_2009&action=edit 26]
[http://2009.igem.org/wiki/index.php?title=July/27_July_2009&action=edit 27] [http://2009.igem.org/wiki/index.php?title=July/28_July_2009&action=edit 28] [http://2009.igem.org/wiki/index.php?title=July/29_July_2009&action=edit 29] [http://2009.igem.org/wiki/index.php?title=July/30_July_2009&action=edit 30] [http://2009.igem.org/wiki/index.php?title=July/31_July_2009&action=edit 31]
August
MTWTFSS
          [http://2009.igem.org/August/1_August_2009 1] [http://2009.igem.org/August/2_August_2009 2]
[http://2009.igem.org/August/3_August_2009 3] [http://2009.igem.org/August/4_August_2009 4] [http://2009.igem.org/August/5_August_2009 5] [http://2009.igem.org/August/6_August_2009 6] [http://2009.igem.org/August/7_August_2009 7] [http://2009.igem.org/August/8_August_2009 8] [http://2009.igem.org/August/9_August_2009 9]
[http://2009.igem.org/August/10_August_2009 10] [http://2009.igem.org/August/11_August_2009 11] [http://2009.igem.org/August/12_August_2009 12] [http://2009.igem.org/August/13_August_2009 13] [http://2009.igem.org/August/14_August_2009 14] [http://2009.igem.org/August/15_August_2009 15] [http://2009.igem.org/August/16_August_2009 16]
[http://2009.igem.org/August/17_August_2009 17] [http://2009.igem.org/August/18_August_2009 18] [http://2009.igem.org/August/19_August_2009 19] [http://2009.igem.org/August/20_August_2009 20] [http://2009.igem.org/August/21_August_2009 21] [http://2009.igem.org/August/22_August_2009 22] [http://2009.igem.org/August/23_August_2009 23]
[http://2009.igem.org/August/24_August_2009 24] [http://2009.igem.org/August/25_August_2009 25] [http://2009.igem.org/August/26_August_2009 26] [http://2009.igem.org/August/27_August_2009 27] [http://2009.igem.org/August/28_August_2009 28] [http://2009.igem.org/August/29_August_2009 29] [http://2009.igem.org/August/30_August_2009 30]
[http://2009.igem.org/August/31_August_2009 31]
September
MTWTFSS
  [http://2009.igem.org/September/1_September_2009 1] [http://2009.igem.org/September/2_September_2009 2] [http://2009.igem.org/September/3_September_2009 3] [http://2009.igem.org/September/4_September_2009 4] [http://2009.igem.org/September/5_September_2009 5] [http://2009.igem.org/September/6_September_2009 6]
[http://2009.igem.org/September/7_September_2009 7] [http://2009.igem.org/September/8_September_2009 8] [http://2009.igem.org/September/9_September_2009 9] [http://2009.igem.org/September/10_September_2009 10] [http://2009.igem.org/September/11_September_2009 11] [http://2009.igem.org/September/12_September_2009 12] [http://2009.igem.org/September/13_September_2009 13]
[http://2009.igem.org/September/14_September_2009 14] [http://2009.igem.org/September/15_September_2009 15] [http://2009.igem.org/September/16_September_2009 16] [http://2009.igem.org/September/17_September_2009 17] [http://2009.igem.org/September/18_September_2009 18] [http://2009.igem.org/September/19_September_2009 19] [http://2009.igem.org/September/20_September_2009 20]
[http://2009.igem.org/September/21_September_2009 21] [http://2009.igem.org/September/22_September_2009 22] [http://2009.igem.org/September/23_September_2009 23] [http://2009.igem.org/September/24_September_2009 24] [http://2009.igem.org/September/25_September_2009 25] [http://2009.igem.org/September/26_September_2009 26] [http://2009.igem.org/September/27_September_2009 27]
[http://2009.igem.org/September/28_September_2009 28] [http://2009.igem.org/September/29_September_2009 29] [http://2009.igem.org/September/30_September_2009 30]
October
MTWTFSS
      [http://2009.igem.org/August/1_October_2009 1] [http://2009.igem.org/August/2_October_2009 2] [http://2009.igem.org/August/3_October_2009 3] [http://2009.igem.org/August/4_October_2009 4]
[http://2009.igem.org/August/5_October_2009 5] [http://2009.igem.org/August/6_October_2009 6] [http://2009.igem.org/August/7_October_2009 7] [http://2009.igem.org/August/8_October_2009 8] [http://2009.igem.org/August/9_October_2009 9] [http://2009.igem.org/August/10_October_2009 10] [http://2009.igem.org/August/11_October_2009 11]
[http://2009.igem.org/August/12_October_2009 12] [http://2009.igem.org/August/13_October_2009 13] [http://2009.igem.org/August/14_October_2009 14] [http://2009.igem.org/August/15_October_2009 15] [http://2009.igem.org/August/16_October_2009 16] [http://2009.igem.org/August/17_October_2009 17] [http://2009.igem.org/August/18_October_2009 18]
[http://2009.igem.org/wiki/index.php?title=August/19_October_2009&action=edit 19] [http://2009.igem.org/August/20_October_2009 20] [http://2009.igem.org/wiki/index.php?title=August/21_October_2009&action=edit 21] [http://2009.igem.org/wiki/index.php?title=August/22_October_2009&action=edit 22] [http://2009.igem.org/wiki/index.php?title=August/23_October_2009&action=edit 23] [http://2009.igem.org/wiki/index.php?title=August/24_October_2009&action=edit 24] [http://2009.igem.org/wiki/index.php?title=August/25_October_2009&action=edit 25]
[http://2009.igem.org/wiki/index.php?title=August/26_October_2009&action=edit 26] [http://2009.igem.org/wiki/index.php?title=August/27_October_2009&action=edit 27] [http://2009.igem.org/wiki/index.php?title=August/28_October_2009&action=edit 28] [http://2009.igem.org/wiki/index.php?title=August/29_October_2009&action=edit 29] [http://2009.igem.org/wiki/index.php?title=August/30_October_2009&action=edit 30] [http://2009.igem.org/wiki/index.php?title=August/31_October_2009&action=edit 31]
November
MTWTFSS
            [http://2009.igem.org/wiki/index.php?title=November/1_November_2009&action=edit 1]
[http://2009.igem.org/wiki/index.php?title=November/2_November_2009&action=edit 2] [http://2009.igem.org/wiki/index.php?title=November/3_November_2009&action=edit 3] [http://2009.igem.org/wiki/index.php?title=November/4_November_2009&action=edit 4] [http://2009.igem.org/wiki/index.php?title=November/5_November_2009&action=edit 5] [http://2009.igem.org/wiki/index.php?title=November/6_November_2009&action=edit 6] [http://2009.igem.org/wiki/index.php?title=November/7_November_2009&action=edit 7] [http://2009.igem.org/wiki/index.php?title=November/8_November_2009&action=edit 8]
[http://2009.igem.org/wiki/index.php?title=November/9_November_2009&action=edit 9] [http://2009.igem.org/wiki/index.php?title=November/10_November_2009&action=edit 10] [http://2009.igem.org/wiki/index.php?title=November/11_November_2009&action=edit 11] [http://2009.igem.org/wiki/index.php?title=November/12_November_2009&action=edit 12] [http://2009.igem.org/wiki/index.php?title=November/13_November_2009&action=edit 13] [http://2009.igem.org/wiki/index.php?title=November/14_November_2009&action=edit 14] [http://2009.igem.org/wiki/index.php?title=November/15_November_2009&action=edit 15]
[http://2009.igem.org/wiki/index.php?title=November/16_November_2009&action=edit 16] [http://2009.igem.org/wiki/index.php?title=November/17_November_2009&action=edit 17] [http://2009.igem.org/wiki/index.php?title=November/18_November_2009&action=edit 18] [http://2009.igem.org/wiki/index.php?title=November/19_November_2009&action=edit 19] [http://2009.igem.org/wiki/index.php?title=November/20_November_2009&action=edit 20] [http://2009.igem.org/wiki/index.php?title=November/21_November_2009&action=edit 21] [http://2009.igem.org/wiki/index.php?title=November/22_November_2009&action=edit 22]
[http://2009.igem.org/wiki/index.php?title=November/23_November_2009&action=edit 23] [http://2009.igem.org/wiki/index.php?title=November/24_November_2009&action=edit 24] [http://2009.igem.org/wiki/index.php?title=November/25_November_2009&action=edit 25] [http://2009.igem.org/wiki/index.php?title=November/26_November_2009&action=edit 26] [http://2009.igem.org/wiki/index.php?title=November/27_November_2009&action=edit 27] [http://2009.igem.org/wiki/index.php?title=November/28_November_2009&action=edit 28] [http://2009.igem.org/wiki/index.php?title=November/29_November_2009&action=edit 29]
[http://2009.igem.org/wiki/index.php?title=November/30_November_2009&action=edit 30]


Registry Parts

All of Osaka's Parts and Devices is here

COLOR


K204033 mOrange

placI

K204041 placI+GFP

K204042 placI+YFP

K204043 placI+CFP

K204044 placI+RFP

K204048 placI+mOrange

pcI

K204058 pcI+YFP

K204059 pcI+CFP

K204062 pcI+mOrange

ptetR

K204052 ptetR+mOrange

Carotenoid

K204064 crtE+crtB

K204065 crtI+crtY

K204068 crtZ


K204076 placI+crtE+crtB

K204077 ptetR+crtE+crtB

K204091 placI+crtE+crtB+crtI+crtY

K204095 placI+crtE+crtB+crtZ

K204096 ptetR+crtE+crtB+crtZ

gradation

K204069 cI repressor

K204086 3O6HSL receiver+RFP+cI repressor


SIGNAL (Quorum Sensing)

AI-1

K204047 LasI generater

C4HSL

K204040 RhlI (generater)

K204031 RhlR (receiver)

K204702 RhlR+GFP

3OH,C14 : 1-HSL

K204025 cinI (generater)

K204032 cinR (receiver)

3O6HSL

K204051 LuxR (receiver)

K204700 LuxR+GFP


MOTILITY

K204018 ptetR+EspE


K204501 RBS+FliH

K204502 ptetR+FliH

K204503 pT7+FliH

K204504 C4HSL receiver +FliH


Lab Note

Wet Lab

Week 1 : 8/2~8/8

Brainstorming , parts planning , prepared competent cell

ransformation and miniprep from MIT plate


week 2 : 8/9~8/15

start assembly parts (K204001 ~ K204006) , transformation and mini prep from MIT


Week 3 : 8/16~8/22

assembly parts (~K204015) , parts planning


Week 4 : 8/23~8/29

assembly parts (~K204021)


Week 5 : 8/30~9/5

assembly parts (~K204030)


Week 6 : 9/6~9/12

assembly parts (~K204046)


Week 7 : 9/13~9/19

assembly parts (~K204050)


Week 8 : 9/20~9/26

assembly parts (~K204056)

TEST : sensitivity of inducer and receptor

Sample : 3O6HSL , C4HSL , AI-1 , 3OH,C14:1-HSL

Result : Not confirm the fluorescence...


Week 9 : 9/27~10/3

assembly parts (~K204066 , X1~X8), color (fluorescence) intensity check


Week 10 : 10/4~10/10

assembly parts (~K204078)

Lux receiver sensitivity test, color (fluorescence) intensity check

sequence check : generator and receiver

3O6HSL , C4HSL , AI-1 , 3OH,C14:1-HSL

Get the parts from Team Chiba. Thanks a lot!!


Week 11 : 10/11~10/17

assembly parts (~K2040104)

sequence check : FliH etc…

send Osaka team parts to MIT

FINISH!

Finally we assembled about 120 parts!!


Dry Lab

Week 5 : 8/30~9/5

Planing model.


Week 6 : 9/6~9/12

Studying programing using Matlab.


Week 7 : 9/13~9/19

Making program in case of single coloney.


Week 8 : 9/20~9/26

Making program in case of more than two colonies

Making movie file

Searching paper written some parameter


Week 9 : 9/27~10/3

Blash up our program

Searching paper written some parameter


Week 10 : 10/4~10/10

Making program of color gradation


Week 11 : 10/11~10/17

Blash up our program


Atelie

Week 8 : 9/20~9/26

Making art works


Week 9 : 9/27~10/3

Making art works


Week 10 : 10/4~10/10

Making art works


Week 11 : 10/11~10/17

Making art works


Retrieved from "http://2009.igem.org/Team:Osaka/NOTES"