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Team:CSU Fort Collins/Notebook/Breakdown/Jul
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| - | <title> | + | <title>Biosensor Notes - June</title> |
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| - | <div id='banner'><center><img src=' | + | <div id='banner'><center><img src='https://static.igem.org/mediawiki/2014/b/b2/Team-CSU_Notebook.jpg' style='width:100%; margin-top:-300px'/><a href='https://2014.igem.org/Main_Page'><img src='https://static.igem.org/mediawiki/2014/d/d7/Sysuchina_igemdeLogo.png' style="width:80px; position:absolute; top: 10px; right: 20px; opacity:0.95;"/></a> |
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<ul> | <ul> | ||
<li><a href='/Team:CSU_Fort_Collins/Members/'><span>Members</span></a></li> | <li><a href='/Team:CSU_Fort_Collins/Members/'><span>Members</span></a></li> | ||
| - | <li class='last'><a href='/Team:CSU_Fort_Collins/ | + | <li><a href='/Team:CSU_Fort_Collins/Sponsors/'><span>Sponsors</span></a></li> |
| + | <li class='last'><a href='/Team:CSU_Fort_Collins/Acknowledgements/'><span>Acknowledgements</span></a></li> | ||
</ul> | </ul> | ||
</li> | </li> | ||
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</ul> | </ul> | ||
</li> | </li> | ||
| - | <li class='has-sub'><a href='/Team:CSU_Fort_Collins/Notebook/Breakdown/'><span>Breakdown</span></a> | + | <li class='has-sub'><a href='/Team:CSU_Fort_Collins/Notebook/Breakdown/Jul'><span>Breakdown</span></a> |
<ul> | <ul> | ||
| - | <li><a href=" | + | <li><a href="/Team:CSU_Fort_Collins/Notebook/Breakdown/Jul"><span>July</span></a></li> |
| - | <li><a href=' | + | <li><a href='/Team:CSU_Fort_Collins/Notebook/Breakdown/Aug'><span>August</span></a></li> |
| - | <li class='last'><a href=" | + | <li class='last'><a href="/Team:CSU_Fort_Collins/Notebook/Breakdown/Sep"><span>September</span></a></li> |
</ul> | </ul> | ||
</li> | </li> | ||
| - | <li class='has-sub'><a href='/Team:CSU_Fort_Collins/Notebook/HVP/'><span>High-Value Product</span></a> | + | <li class='has-sub'><a href='/Team:CSU_Fort_Collins/Notebook/HVP/Jun'><span>High-Value Product</span></a> |
<ul> | <ul> | ||
| - | <li><a href=" | + | <li><a href="/Team:CSU_Fort_Collins/Notebook/HVP/Jun"><span>June</span></a></li> |
| - | + | <li class='last'><a href="/Team:CSU_Fort_Collins/Notebook/HVP/Jul"><span>July</span></a></li> | |
| - | + | ||
| - | <li class='last'><a href=" | + | |
</ul> | </ul> | ||
</li> | </li> | ||
| - | <li li class='has-sub'><a href='/Team:CSU_Fort_Collins/Notebook/KillSwitch/'><span>Kill Switch</span></a> | + | <li li class='has-sub'><a href='/Team:CSU_Fort_Collins/Notebook/KillSwitch/Jul'><span>Kill Switch</span></a> |
<ul> | <ul> | ||
| - | <li><a href=" | + | <li><a href="/Team:CSU_Fort_Collins/Notebook/KillSwitch/Jul"><span>July</span></a></li> |
| - | + | <li class='last'><a href="/Team:CSU_Fort_Collins/Notebook/KillSwitch/Sep"><span>September</span></a></li> | |
| - | <li class='last'><a href=" | + | |
</ul> | </ul> | ||
</li> | </li> | ||
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</ul> | </ul> | ||
</li> | </li> | ||
| - | <li class='has-sub'><a href='/Team:CSU_Fort_Collins/ | + | <li class='has-sub'><a href='/Team:CSU_Fort_Collins/Collab/'><span>Human Practices</span></a> |
<ul> | <ul> | ||
<li><a href='/Team:CSU_Fort_Collins/Collab/'><span>Collaboration</span></a></li> | <li><a href='/Team:CSU_Fort_Collins/Collab/'><span>Collaboration</span></a></li> | ||
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</ul> | </ul> | ||
</li> | </li> | ||
| - | <li class='has-sub'><a href='/Team:CSU_Fort_Collins/ | + | <li class='has-sub'><a href='/Team:CSU_Fort_Collins/Parts/'><span>Achievements</span></a> |
<ul> | <ul> | ||
<li><a href='/Team:CSU_Fort_Collins/Parts/'><span>Parts</span></a></li> | <li><a href='/Team:CSU_Fort_Collins/Parts/'><span>Parts</span></a></li> | ||
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</ul> | </ul> | ||
</li> | </li> | ||
| - | <li><a href='/Team:CSU_Fort_Collins/Safety/'><span>Safety</span></a></li> | + | <li class='last'><a href='/Team:CSU_Fort_Collins/Safety/'><span>Safety</span></a></li> |
</ul> | </ul> | ||
</div> | </div> | ||
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<div class="month-group"> | <div class="month-group"> | ||
<a href='/Team:CSU_Fort_Collins/Notebook/Breakdown/Jul'>July</a> | <a href='/Team:CSU_Fort_Collins/Notebook/Breakdown/Jul'>July</a> | ||
| + | </div> | ||
<p>Lac Promoter and FadD Assembly</p> | <p>Lac Promoter and FadD Assembly</p> | ||
| - | |||
<div class="month-group"> | <div class="month-group"> | ||
<a href='/Team:CSU_Fort_Collins/Notebook/Breakdown/Aug'>August</a> | <a href='/Team:CSU_Fort_Collins/Notebook/Breakdown/Aug'>August</a> | ||
| - | <p> | + | </div> |
| - | + | <p>More of the Same, But Better</p> | |
<div class="month-group"> | <div class="month-group"> | ||
<a href='/Team:CSU_Fort_Collins/Notebook/Breakdown/Sep'>September</a> | <a href='/Team:CSU_Fort_Collins/Notebook/Breakdown/Sep'>September</a> | ||
| - | + | </div> | |
| - | + | <p>One Thousand and One Nights</p> | |
</div> | </div> | ||
<div class='page'> | <div class='page'> | ||
| - | <p> | + | <p> <h2>JULY</h2> |
| - | + | ||
<h4>Tuesday, July 1 - Wednesday, July 16</h4> | <h4>Tuesday, July 1 - Wednesday, July 16</h4> | ||
Researched which promoters, primers, sequences, and assembly techniques to use and developed a plan of action. | Researched which promoters, primers, sequences, and assembly techniques to use and developed a plan of action. | ||
<h4>Thursday, July 17</h4> | <h4>Thursday, July 17</h4> | ||
| - | Designed primers to connect biobrick lab promoter and the FadD gene from E. coli. | + | Designed primers to connect biobrick lab promoter and the FadD gene from <u>E. coli</u>. |
<h4>Friday, July 18</h4> | <h4>Friday, July 18</h4> | ||
| - | With competent cells, used the transformation protocol to add our two different lac promoters to E. coli and grew overnight at 37 | + | With competent cells, used the transformation protocol to add our two different lac promoters to <u>E. coli</u> and grew overnight at 37 °C. Used lac promoters from the biobrick distribution kit (2013 iGEM Distribution Kit, Plate 5, Well 1D (ampicillin resistance) and 2014 iGEM Distribution Kit, Plate 3, Well 4G (chloramphenicol resistance)). |
<h4>Sunday, July 19</h4> | <h4>Sunday, July 19</h4> | ||
| - | Removed plates from the incubator and stored at 4 | + | Removed plates from the incubator and stored at 4 °C. |
<h4>Monday, July 21</h4> | <h4>Monday, July 21</h4> | ||
| - | The plate using the promoter from 1D had no growth. The plate with the promoter from 4G had two small colonies. We replated the colonies using the rest of our transformed competent cells. Incubated again overnight at 37 | + | The plate using the promoter from 1D had no growth. The plate with the promoter from 4G had two small colonies. We replated the colonies using the rest of our transformed competent cells. Incubated again overnight at 37 °C. |
<h4>Tuesday, July 22</h4> | <h4>Tuesday, July 22</h4> | ||
| - | The plate using the promoter from 1D still had no growth. The plate with the promoter from 4G had a lawn. Streaked the 4G plate to create individual colonies. Incubated overnight at 37 | + | The plate using the promoter from 1D still had no growth. The plate with the promoter from 4G had a lawn. Streaked the 4G plate to create individual colonies. Incubated overnight at 37 °C. |
<h4>Wednesday, July 23</h4> | <h4>Wednesday, July 23</h4> | ||
| - | The 4G plate had good growth with many individual colonies. We created two 2 mL overnight cultures using 1 mL LB/1 | + | The 4G plate had good growth with many individual colonies. We created two 2 mL overnight cultures using 1 mL LB/1 μL CM and a colony from our streaked plate. Incubated overnight at 37 °C and 225 rpm. |
<h4>Thursday, July 24</h4> | <h4>Thursday, July 24</h4> | ||
| - | Removed the tubes from the incubator and stored at 4 | + | Removed the tubes from the incubator and stored at 4 °C. |
<h4>Friday, July 25</h4> | <h4>Friday, July 25</h4> | ||
| - | The primers for FadD gene came in. Created a 100 | + | The primers for FadD gene came in. Created a 100 μM stock solution. Diluted by 10X to make a working solution. Following the PCR protocol outlined in the Cloning a Gene into a Plasmid protocol, PCRed 4 replicates. Used a 68 °C annealing temperature and a 1 minute 30 seconds extension. Stored the PCR product tubes at 4 °C. |
| - | Miniprepped 1 mL of overnight cultures to extract the DNA containing the desired lac promoter as directed in the Miniprep Protocol. The products of the miniprep were stored at -20 | + | Miniprepped 1 mL of overnight cultures to extract the DNA containing the desired lac promoter as directed in the <a href='/Team:CSU_Fort_Collins/Notebook/Protocols=Miniprep'>Miniprep Protocol</a>. The products of the miniprep were stored at -20 °C. |
Created 2 more overnight cultures using the same protocol as before, this time seeding them using what was left of the previous overnight cultures. | Created 2 more overnight cultures using the same protocol as before, this time seeding them using what was left of the previous overnight cultures. | ||
<h4>Tuesday, July 29</h4> | <h4>Tuesday, July 29</h4> | ||
| - | Used two <a href='Team:CSU_Fort_Collins/Notebook/Protocols=Gel'>gel electrophoresis</a> protocols to determine if the PCR worked. Both gels failed. | + | Used two <a href='/Team:CSU_Fort_Collins/Notebook/Protocols=Gel'>gel electrophoresis</a> protocols to determine if the PCR worked. Both gels failed. |
<h4>Wednesday, July 30</h4> | <h4>Wednesday, July 30</h4> | ||
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<br><br> | <br><br> | ||
| - | <center><a href='/Team:CSU_Fort_Collins/Notebook/ | + | <center><a href='/Team:CSU_Fort_Collins/Notebook/Biosensor/Aug' id='navi' style='margin-left:-20px; margin-right:40px'> Previous</a> <a href='/Team:CSU_Fort_Collins/Notebook/Breakdown/Aug' id='navi'>Next </a></center> |
</div> | </div> | ||
Latest revision as of 20:15, 17 October 2014
Breakdown Daily Notes
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Lac Promoter and FadD Assembly
More of the Same, But Better
One Thousand and One Nights
