Team:Chiba/Calendar-Home/12 September 2008

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==Laboratory work==
==Laboratory work==
===Team:Input===
===Team:Input===
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*Pbad/araCのL-アラビノースによるスイッチングテスト
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*Pbad/araC:L-Arabinose induction test
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I1304(Ptet-RBS-GFP,XL10G)×1()(ネガティブコントロール用),R0051(PcI)×4(②~⑤)のコロニーをつついて2ml培養。
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*[http://partsregistry.org/Part:BBa_I1304 BBa_I1304]×1(1)(negative control)
 +
*[http://partsregistry.org/Part:BBa_R0051 BBa_R0051]×4(1)~(5)Select single colony from Agar Plate and introduce to 2mlLB each other.
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37℃,12h後液が濁っていたら、ODをはかり、48穴deep wellに①~⑤の菌液をそれぞれ190μl入れた。
 
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#incubated cultures at 37 &degC; for 12hours.
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#Measured OD<sub>600</sub>.
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L-アラビノースを以下のような濃度になるように入れた。
 
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混ぜ表
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Reaction mixtures were mixed in 48-well plate(deep well).
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<table width="100" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
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<td width="257"></td>
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<td>20%L-アラビノース</td><td>濃度</td>
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<table width="257" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
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<td width="257">Sample No.</td>
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<td>1</td><td>2</td><td>3</td><td>4</td><td>5</td>
<tr>
<tr>
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<td> </td>
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<td>Cultures(&mu;L)</td>
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<td>0μl</td><td>0%(N.C)</td>
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<td>0&mu;l</td><td>1.9&mu;l</td><td>19&mu;l</td><td>190&mu;l</td><td>-</td>
</tr>
</tr>
<tr>
<tr>
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<td> </td>
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<td> 20%L-Arabinose(&mu;L)</td>
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<td>1.9μl</td><td>0.2%</td>
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<td>0&mu;l</td><td>1.9&mu;l</td><td>19&mu;l</td><td>190&mu;l</td><td>-</td>
</tr>
</tr>
<tr>
<tr>
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<td> </td>
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<td>L-Arabinase conc.</td>
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<td>19μl</td><td>2%</td>
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<td>0%(N.C)</td><td>0.2%</td><td>2%</td><td>20%</td><td>P.C</td>
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</tr>
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<tr>
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<td> ④</td>
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<td>190μl</td><td>20%</td>
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</tr>
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<tr>
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<td> ⑤</td>
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<td>-</td><td>P.C</td>
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</tr>
</tr>
</table>
</table>
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L-アラビノースを入れてから15min,1h,2h,6h,12h,24h後に蛍光強度を計測した。
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Measured the Green Fluorescence intensity by Fluoroskan Ascent 2.5(program:Ascent Software Version 2.6).
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Add L-arabinose and 30min,1h,2h,6h,12h,24h later, we measured intensicty of fluorescence.
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結果は以下のとおり。
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Result:
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<table width="400" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
<table width="400" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
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<td width="257"></td>
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<td width="2300">Time after arabinose induction  / Sample No.</td>
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<td></td><td></td><td></td><td></td><td></td>
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<td>1</td><td>2</td><td>3</td><td>4</td><td>5</td>
<tr>
<tr>
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<td>L-アラビノース入れて30分後</td>
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<td>30min</td>
<td>8.890</td><td>9.752</td><td>10.10</td><td>9.413</td><td>9.701</td>
<td>8.890</td><td>9.752</td><td>10.10</td><td>9.413</td><td>9.701</td>
</tr>
</tr>
<tr>
<tr>
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<td>L-アラビノース入れて1h後</td>
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<td>1h</td>
<td>9.161</td><td>13.93</td><td>17.37</td><td>14.97</td><td>10.60</td>
<td>9.161</td><td>13.93</td><td>17.37</td><td>14.97</td><td>10.60</td>
</tr>
</tr>
<tr>
<tr>
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<td>L-アラビノース入れて2h後</td>
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<td>2h</td>
<td>9.417</td><td>27.96</td><td>40.25</td><td>29.87</td><td>10.72</td>
<td>9.417</td><td>27.96</td><td>40.25</td><td>29.87</td><td>10.72</td>
</tr>
</tr>
<tr>
<tr>
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<td>L-アラビノース入れて6h後</td>
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<td>6h</td>
<td>9.933</td><td>74.99</td><td>150.9</td><td>125.0</td><td>10.04</td>
<td>9.933</td><td>74.99</td><td>150.9</td><td>125.0</td><td>10.04</td>
<tr>
<tr>
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<td>L-アラビノース入れて12h後</td>
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<td>12h</td>
<td>10.64</td><td>99.39</td><td>202.88</td><td>72.08</td><td>11.76</td>
<td>10.64</td><td>99.39</td><td>202.88</td><td>72.08</td><td>11.76</td>
</tr>
</tr>
<tr>
<tr>
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<td>L-アラビノース入れて24h後</td>
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<td>24h</td>
<td>14.66</td><td>105.6</td><td>447.3</td><td>33.31</td><td>17.11</td>
<td>14.66</td><td>105.6</td><td>447.3</td><td>33.31</td><td>17.11</td>
</tr>
</tr>
</table>
</table>
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===Team:Communication===
===Team:Communication===

Latest revision as of 21:26, 29 October 2008

>Home | Notebook

11 September 2008 <|> 13 September 2008

Contents

Laboratory work

Team:Input

  • Pbad/araC:L-Arabinose induction test
  • [http://partsregistry.org/Part:BBa_I1304 BBa_I1304]×1(1)(negative control)
  • [http://partsregistry.org/Part:BBa_R0051 BBa_R0051]×4(1)~(5)Select single colony from Agar Plate and introduce to 2mlLB each other.


  1. incubated cultures at 37 &degC; for 12hours.
  2. Measured OD600.


Reaction mixtures were mixed in 48-well plate(deep well).


Sample No. 12345
Cultures(μL) 0μl1.9μl19μl190μl-
20%L-Arabinose(μL) 0μl1.9μl19μl190μl-
L-Arabinase conc. 0%(N.C)0.2%2%20%P.C

Measured the Green Fluorescence intensity by Fluoroskan Ascent 2.5(program:Ascent Software Version 2.6). Add L-arabinose and 30min,1h,2h,6h,12h,24h later, we measured intensicty of fluorescence. Result:

Time after arabinose induction / Sample No. 12345
30min 8.8909.75210.109.4139.701
1h 9.16113.9317.3714.9710.60
2h 9.41727.9640.2529.8710.72
6h 9.93374.99150.9125.010.04
12h 10.6499.39202.8872.0811.76
24h 14.66105.6447.333.3117.11

Team:Communication

no work

Team:Output

no work