Team:CSU Fort Collins/Notebook/HVP/Jun
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- | + | <li class='last'><a href="/Team:CSU_Fort_Collins/Notebook/HVP/Jul"><span>July</span></a></li> | |
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- | <center><h1> | + | <center><h1>High-Value Product Daily Notes</h1> |
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<a href='/Team:CSU_Fort_Collins/Notebook/Breakdown/Jun'>Jun</a> | <a href='/Team:CSU_Fort_Collins/Notebook/Breakdown/Jun'>Jun</a> | ||
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<a href='/Team:CSU_Fort_Collins/Notebook/Breakdown/Jul'>July</a> | <a href='/Team:CSU_Fort_Collins/Notebook/Breakdown/Jul'>July</a> | ||
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<p>pMBI and pMevT Assembly</p> | <p>pMBI and pMevT Assembly</p> | ||
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- | <h2> | + | <h2>JUNE</h2> |
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- | <h4> | + | <h4>Monday, June 2 - Friday, June 6</h4> |
- | + | Researched potential products and decided on the addition of the mevalonate pathway in order to direct <u>E. coli</u> to terpenoid production. | |
- | <h4> | + | <h4>Monday, June 9 - Monday, June 16</h4> |
- | + | Researched the mevalonate pathway in <u>E. coli</u> and sequenced the genes to be inserted into puc19. Designed primers as specified in the <a href='/Team:CSU_Fort_Collins/Notebook/Protocols=Gibson'>Gibson Assembly Protocol</a> for transformation of the mevalonate pathway from yeast into <u>E. coli</u>. | |
- | + | <h4>Thursday, June 19</h4> | |
+ | Ran a <a href='Team:CSU_Fort_Collins/Notebook/Protocol/Gel'>gel electrophoresis</a> to try and confirm presence of DNA in obtained yeast samples. No results. | ||
- | + | <h4>Tuesday, June 24</h4> | |
+ | Mixed and autoclaved two 500 mL bottles of LB Broth with Agar and ampicillin. Poured two sleeves of plates to use for culturing. | ||
- | <h4> | + | <h4>Wednesday, June 25</h4> |
- | + | Took a small sample of <u>E. coli</u> with puc19 from a previous colony and suspended in LB Broth. Shook at 225 rpm in incubator at 37° C overnight to culture. | |
- | <h4> | + | <h4>Thursday, June 26</h4> |
- | + | Completed a miniprep of <u>E. coli</u> with puc19 from the overnight culture. Followed <a href='/Team:CSU_Fort_Collins/Notebook/Protocol/Miniprep'>plasmid miniprep protocol</a> as given in the Informatics (Life Technologies) kit. | |
- | <h4> | + | <h4>Friday, June 27</h4> |
- | + | Completed PCR with the 5 pieces according to the mixture of New England Biotechnologies' Q5 2X Master Mix protocol. Ran a gel electrophoresis to confirm success of PCR. Amplification of PMK unsuccessful. Amplification of all other portions of the plasmid successful. | |
+ | <h4>Monday, June 30</h4> | ||
+ | Modified the PCR thermalcycler program (but used the same 50 μL mixture) and ran a gel to check results. All attempts unsuccessful - suspected due to lack of presence of SYBR green in initial mixture. | ||
</p> | </p> | ||
<br><br> | <br><br> | ||
- | <center><a href='/Team:CSU_Fort_Collins/Notebook/ | + | <center><a href='/Team:CSU_Fort_Collins/Notebook/Breakdown/Aug' id='navi' style='margin-left:-20px; margin-right:40px'> Previous</a> <a href='/Team:CSU_Fort_Collins/Notebook/HVP/Jul' id='navi'>Next </a></center> |
</div> | </div> | ||
Latest revision as of 20:17, 17 October 2014
High-Value Product Daily Notes
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