Team:Duke/Notebook

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<p class="big"> Lab Notebook </p>
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<div class="month"><a href="https://2014.igem.org/Team:Duke/Notebook/Overview">Overview of Each Month </a></div>
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<div class="month"><a href="https://2014.igem.org/Team:Duke/Notebook/Months">Notebook Entries by Month</a></div>
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<div class="month"><a href="https://2014.igem.org/Team:Duke/Notebook/Protocols">Protocols</a></div>
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<td colspan="7"><a href="https://2014.igem.org/Team:Duke/Notebook#apr"> April 2014 </a></td>
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<td colspan="7"><i> Month 1 of Project </i></td>
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<td>sun</td>
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<td>1</td>
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<td>2</td>
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<td>3</td>
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<td>4</td>
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<td>5</td>
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<td>6</td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#apr7">7</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#apr8">8</a></td>
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<td>9</td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#apr10">10</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#apr11">11</a></td>
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<td>12</td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#apr13">13</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#apr14">14</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#apr15">15</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#apr16">16</a></td>
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<td>17</td>
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<td>18</td>
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<td>19</td>
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<td>20</td>
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<td>21</td>
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<td>22</td>
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<td>23</td>
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<td>27</td>
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<td colspan="7"> May 2014 </td>
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<td colspan="7" align=center><i> Month 2 of Project </i></td>
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<td>sun</td>
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<td>mon</td>
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<td>1</td>
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<td>3</td>
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<td>4</td>
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<td>5</td>
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<td>6</td>
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<td>7</td>
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<td>8</td>
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<td>9</td>
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<td>10</td>
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<td>11</td>
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<td>12</td>
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<td>13</td>
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<td>14</td>
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<td>15</td>
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<td>16</td>
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<td>17</td>
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<td>18</td>
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<td>19</td>
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<td>20</td>
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<td>21</td>
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<td>22</td>
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<td>23</td>
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<td>24</td>
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<td>25</td>
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<td>26</td>
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<td>27</td>
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<td>28</td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#may29">29</a></td>
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<td>30</td>
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<td>31</td>
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<td colspan="7"> June 2014 </td>
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<td colspan="7" align=center><i> Month 3 of Project </i></td>
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<td>sun</td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jun1">1</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jun2">2</a></td>
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<td>3</td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jun4">4</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jun5">5</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jun6">6</a></td>
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<td>7</td>
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<td>8</td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jun9">9</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jun10">10</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jun11">11</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jun12">12</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jun13">13</a></td>
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<td>14</td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jun15">15</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jun16">16</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jun17">17</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jun18">18</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jun19">19</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jun20">20</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jun21">21</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jun22">22</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jun23">23</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jun24">24</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jun25">25</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jun26">26</a></td>
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<td>27</td>
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<td>28</td>
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<td>29</td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jun30">30</a></td>
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<td colspan="7"> July 2014 </td>
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<td colspan="7" align=center><i> Month 4 of Project </i></td>
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<td>sun</td>
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<td>mon</td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jul1">1</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jul2">2</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jul3">3</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jul4">4</a></td>
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<td ><a href="https://2014.igem.org/Team:Duke/Notebook#jul5">5</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jul6">6</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jul7">7</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jul8">8</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jul9">9</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jul10">10</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jul11">11</a></td>
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<td>12</td>
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<td>13</td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jul14">14</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jul15">15</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jul16">16</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jul17">17</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jul18">18</a></td>
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<td>19</td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jul20">20</a></td>
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<td><a href="https://2014.igem.org/Team:Duke/Notebook#jul21">21</a></td>
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<td>22</td>
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<td colspan="7"> August 2014 </td>
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<td colspan="7" align=center><i> Month 5 of Project </i></td>
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<td align=center>sun</td>
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<td align=center>mon</td>
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<td align=center>1</td>
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<td align=center>3</td>
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<td align=center>10</td>
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<td align=center>11</td>
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<td align=center>17</td>
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<td align=center>18</td>
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<td align=center>24</td>
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<td align=center>25</td>
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<td align=center>31</td>
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<td colspan="7"> September 2014 </td>
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<td colspan="7" align=center><i> Month 6 of Project </i></td>
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<td align=center>sun</td>
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<td align=center>1</td>
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<td align=center>7</td>
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<td align=center>14</td>
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<td align=center>21</td>
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<td align=center>22</td>
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<td align=center>28</td>
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<td colspan="7"> October 2014 </td>
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<td colspan="7" align=center><i> Month 7 of Project </i></td>
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<td align=center>sun</td>
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<td align=center>mon</td>
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<td align=center>tue</td>
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<td align=center>thu</td>
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<td align=center>1</td>
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<td align=center>2</td>
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<td align=center>3</td>
+
-
<td align=center>4</td>
+
-
</tr>
+
-
<tr>
+
-
<td align=center>5</td>
+
-
<td align=center>6</td>
+
-
<td align=center>7</td>
+
-
<td align=center>8</td>
+
-
<td align=center>9</td>
+
-
<td align=center>10</td>
+
-
<td align=center>11</td>
+
-
</tr>
+
-
<tr>
+
-
<td align=center>12</td>
+
-
<td align=center>13</td>
+
-
<td align=center>14</td>
+
-
<td align=center>15</td>
+
-
<td align=center>16</td>
+
-
<td align=center>17</td>
+
-
<td align=center>18</td>
+
-
</tr>
+
-
<tr>
+
-
<td align=center>19</td>
+
-
<td align=center>20</td>
+
-
<td align=center>21</td>
+
-
<td align=center>22</td>
+
-
<td align=center>23</td>
+
-
<td align=center>24</td>
+
-
<td align=center>25</td>
+
-
</tr>
+
-
<tr>
+
-
<td align=center>26</td>
+
-
<td align=center>27</td>
+
-
<td align=center>28</td>
+
-
<td align=center>29</td>
+
-
<td align=center>30</td>
+
-
<td align=center>31</td>
+
-
<td align=center></td>
+
-
</tr>
+
-
<tr>
+
-
<td align=center></td>
+
-
<td align=center></td>
+
-
<td align=center></td>
+
-
<td align=center></td>
+
-
<td align=center></td>
+
-
<td align=center></td>
+
-
<td align=center></td>
+
-
</tr>
+
-
</table>
+
-
</div>
+
-
</div>
+
-
 
+
-
<div id="rightnb">
+
-
 
+
-
<div class="day">
+
-
<a id="apr"><h1>April Overview</h1></a>
+
-
<div class="obj">Objectives</div>
+
-
<div class="lab">
+
-
<p>This is what we were trying to accomplish in April. </p>
+
-
</div>
+
-
</div>
+
-
 
+
-
 
+
-
 
+
-
 
+
-
<div class="day">
+
-
<a id="apr"><h1>May, June Overview</h1></a>
+
-
<div class="obj">Objectives</div>
+
-
<div class="lab">
+
-
<p>This is what we were trying to accomplish in May and June. </p>
+
-
</div>
+
-
</div>
+
-
 
+
-
<div class="day">
+
-
<a id="may29"><h2>May 29</h2></a>
+
-
<div class="obj">Objective: Prepare Chemically competent E. Coli</div>
+
-
<div class="ppl">Matthew Faw</div>
+
-
<div class="lab">
+
-
<p> Followed Charlie’s Cloning Protocols to prepare chemically competent E. Coli </p>
+
-
<ul>
+
-
<li> Once E. Coli were prepared, the solution was aliquoted into 12 labeled tubes (450l in each tube) and tubes were stored in iGEM box in -80C cooler on far left</li>
+
-
</ul>
+
-
<p>Next Steps: </p>
+
-
<ul>
+
-
<li> Transformation </li>
+
-
</ul>
+
-
</div>
+
-
</div>
+
-
 
+
-
 
+
-
<div class="day">
+
-
<a id="jun1"><h2>June 1</h2></a>
+
-
<div class="obj">Objective: Transform Chemically competent E. Coli (CCEC)</div>
+
-
<div class="ppl">Matthew Faw</div>
+
-
<div class="lab">
+
-
<p> Transformation of CCEC with RFP Construct of varying concentrations+control with no DNA</p>
+
-
<ul>
+
-
<li>Followed Charlie’s Cloning Protocols</li>
+
-
<ul>
+
-
<li>Mistakenly used all of DNA construct from the kit… So not sure if the transformation will be successful</li>
+
-
</ul>
+
-
<li>Plated the transformed DNA and put in incubator at 37C</li>
+
-
<li>Results (6/2/14)--Transformation unsuccessful because improper procedure was done by MFaw</li>
+
-
</ul>
+
-
 
+
-
<p> Next Steps:</p>
+
-
<ul>
+
-
<li>Look at plates, compare cultures</li>
+
-
</ul>
+
-
</div>
+
-
</div>
+
-
 
+
-
<div class="day">
+
-
<a id="jun2"><h2>June 2</h2></a>
+
-
<div class="obj">Objective: Redo yesterday’s transformation</div>
+
-
<div class="ppl">Matthew Faw</div>
+
-
<div class="lab">
+
-
<p> Due to my failure to correctly conduct the transformation yesterday, it was necessary to redo the transformation.</p>
+
-
<ul>
+
-
<li>Followed Charlie’s Cloning Protocols</li>
+
-
<ul><li>Added 1 ul of .5, 5, 10, 20, 50, and 0 (control) pg/lof RFP Construct to chemically competent E. Coli (CCEC), and followed procedure</li></ul>
+
-
<li>Plated the transformed DNA on 6 separate plates, put in 37C overnight</li>
+
-
<li>Results: (6/3/14)-Transformation unsuccessful. Try the transformation again with Charlie’s cells and with new cells grown using iGEM’s protocol</li>
+
-
</ul>
+
-
 
+
-
<p>Next Steps:</p>
+
-
<ul>
+
-
<li>Look at plates, compare cultures, etc. </li>
+
-
</ul>
+
-
</div>
+
-
</div>
+
-
 
+
-
 
+
-
 
+
-
6/4/14
+
-
 
+
-
Objective: Prepare buffers and mediums for new CCEC protocol
+
-
Matthew Farnitano, TJ Ciesla, Mike Zhu, Charlie Cooper
+
-
Prepare SOB Medium for bacterial transformation
+
-
Protocol from iGEM’s parts website http://parts.igem.org/Help:Protocols/Competent_Cells
+
-
Made 1 L, autoclaved and stored in two 500 mL containers in cold room
+
-
medium still appeared cloudy before autoclave--may just be new recipe
+
-
Prepare CCMB 80 Buffer for making chemically competent E. coli cells
+
-
Protocol from iGEM’s parts website http://parts.igem.org/Help:Protocols/Competent_Cells
+
-
Made 1 L, filtered and stored in two 500 mL containers in cold room
+
-
pH 6.34 (overshot a few times pH adjustment, but no noticeable precipitate formed
+
-
Autoclave two 500 mL culture flasks
+
-
For CCEC protocol
+
-
With water inside to remove detergent residues
+
-
Next steps:
+
-
Prepare CCEC
+
-
 
+
-
Objective: Attempt to grow some transformed cell cultures with Charlie’s CCEC
+
-
Matthew Faw
+
-
The second (and more properly conducted) attempt to transform the CCEC with RFP construct BBa_J04450 from 6/2 failed.  Today, we are trying to see if we can get any transformed cells to grow in plates. 
+
-
Followed Charlie’s Cloning protocols,with slight modifications
+
-
Added 5 lof 0 (control) and 50pg/lRFP Construct BBa_J04450 to Charlie’s chemically competent E. Coli, and followed procedure
+
-
Plated the transformed DNA on 2 separate plates, put in put in 37C overnight
+
-
Results: No colonies grew (6/5/14)
+
-
Next Steps:
+
-
-Examine plates to see if any cultures grew
+
-
-Attempt the transformation with the cell cultures growns using iGEM’s standard protocols that can be found here: http://parts.igem.org/Help:Protocols/Competent_Cells
+
-
-Lab currently in the process of making these CCEC
+
-
 
+
-
Objective: Prepare pdCas9 marrafini and pCsy4 Arkin plasmids to be miniprepped
+
-
Matthew Faw, Charlie Cooper
+
-
Prepare pdCas9 and pCsy4 to be miniprepped tomorrow
+
-
Put pdCas9 into a culture tube with 5ml SOC+Chloramphenicol
+
-
Put pCsy4 in a culture tube with 5ml SOC+Amp
+
-
Left both to shake at 37C to grow the plasmid DNA--Charlie will retrieve them
+
-
 
+
-
Next steps:
+
-
Miniprep plasmid DNA
+
-
 
+
-
 
+
-
<div class="day">
+
-
<a id=""><h2></h2></a>
+
-
<div class="obj"></div>
+
-
<div class="ppl"></div>
+
-
<div class="lab">
+
-
<p></p>
+
-
<ul>
+
-
<li></li>
+
-
</ul>
+
-
</div>
+
-
</div>
+
-
 
+
-
 
+
-
</div>
+
</body>
</body>
</html>
</html>

Latest revision as of 02:14, 18 October 2014

Overview of Each Month
Notebook Entries by Month
Protocols

Retrieved from "http://2014.igem.org/Team:Duke/Notebook"