<p>In organic extractions, many of our experiments failed due to improper procedure. We found the following to help with successful extraction: ensure an emulsion has not formed (to remove, wait a long time or centrifuge the mixture); use compatible materials (ensure containers, tubes, tips, filters, and other materials are DMSO-safe and do not degrade with the specific organic solvent being used, and also check for organic solvent miscibility); ensure product purity by doing pre-extraction wash step. This pre-extraction wash step is specific to LC-MS detection, as it is often more sensitive and requires higher purity compared to HPLC. Our protocol for this pre-extraction wash step can be found on our experiments tab.</p>
<p>We also encountered issues with cell growth on selective agar plates, indicating problems with electroporation. To resolve these issues, we employed the following strategies: use of fresh electrocompetent cells, increase of the recovery time, increase of the incubation time, increase of the amount of DNA added. In a few of our electroporation attempts, we encountered arcing, due to insufficient purification of added DNA. To circumvent this problem, we often doubled the amount of DNA mixture undergoing drop dialysis for purification in order to have a backup purified DNA stock to immediately electroporate again with new cells in the event of arcing.</p>
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<h2>iGEM Parts Application Summary</h2>
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<h3>Bronze Medal Basic Part Application</h3>
<p>We have submitted a basic part for consideration, using the specifications of GmCHR5 or <ahref="https://parts.igem.org/Part:BBa_K4947024"target="_blank">BBa_K4947024</a>.</p>
<h3>Silver Medal Composite Part Application</h3>
<p>Our team is presenting a composite part for the Silver medal, integrating components from A111 or <ahref="https://parts.igem.org/Part:BBa_K4947034"target="_blank">BBa_K4947034</a>. This composite part is not only our submission for the Silver but is also vying for the special prize targeting the Gold medal category.</p>
<h3>Contributions Overview</h3>
<p>For transparency and documentation, we have also listed our extensive contributions to the parts registry, comprising 45 parts (ranging from BBa_K4947000 to BBa_K4947044). </p>
<p>Find below a brief overview of the literature we reviewed as a part of our decision-making process.</p>
<iframeid="cont-pdf"src="https://static.igem.wiki/teams/4947/wiki/lit-review-on-extraction-malonyl-coa-p-coumaric-acid-dzn-optimization-daidzein-optimizations.pdf">Find above a brief overview of the literature we reviewed as a part of our decision-making process.</iframe>
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<h2>iGEM Parts Submission Summary</h2>
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<p>We submitted our extensive contributions to the parts registry, comprising 45 parts (ranging from BBa_K4947000 to BBa_K4947044). </p>
<h3>Silver Medal and Special Prize Composite Part Application</h3>
<p>Our team is presenting a composite part for the Silver medal, which is A111 or <ahref="https://parts.igem.org/Part:BBa_K4947034"target="_blank">BBa_K4947034</a> in the registry.</p>
