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<h2 id="vbid-7d38c6a1-8ye3bio7" class="preview-element preview-title magic-circle-holder inner-page text-element quick-text-style-menu allow-mobile-hide" data-menu-name="PREVIEW_TITLE" >usMA-WEST POINT<br></h2> | <h2 id="vbid-7d38c6a1-8ye3bio7" class="preview-element preview-title magic-circle-holder inner-page text-element quick-text-style-menu allow-mobile-hide" data-menu-name="PREVIEW_TITLE" >usMA-WEST POINT<br></h2> | ||
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− | <ol><li class="font_7" style="text-align: justify;">OR vectors were purchased from OriGene and | + | <ol><li class="font_7" style="text-align: justify;">OR vectors were purchased from OriGene and used to transform XL-10 Gold Ultracompetent cells by heat shocking at 42 °C<br></li><li class="font_7" style="text-align: justify;">Cells were cultured in LB broth to amplify vectors<br></li><li class="font_7" style="text-align: justify;">DNA gels were run with specific restrion enzymes to determine multiple cloaning sites<br></li><li class="font_7" style="text-align: justify;">Relevant documents<br><ol><li class="font_7" style="text-align: justify;"><a data-cke-saved-href="https://www.mcdb.ucla.edu/Research/Banerjee/protocols/maxiprep--Qiagen.pdf" href="https://www.mcdb.ucla.edu/Research/Banerjee/protocols/maxiprep--Qiagen.pdf">Qiagen MaxiPrep </a><br></li><li class="font_7" style="text-align: justify;"><a data-cke-saved-href="http://www.chem-agilent.com/pdf/strata/200314.pdf" href="http://www.chem-agilent.com/pdf/strata/200314.pdf">XL10-Gold Ultracompetent cells</a><br></li><li class="font_7" style="text-align: justify;"><a data-cke-saved-href="https://tools.thermofisher.com/content/sfs/manuals/Lipofectamine_2000_Reag_protocol.pdf" href="https://tools.thermofisher.com/content/sfs/manuals/Lipofectamine_2000_Reag_protocol.pdf">Lipofectamine 2000 Reagent</a><br></li><li class="font_7" style="text-align: justify;"><a data-cke-saved-href="https://www.neb.com/tools-and-resources/usage-guidelines/cloning-guide" href="https://www.neb.com/tools-and-resources/usage-guidelines/cloning-guide">DNA cloning</a><br></li><li class="font_7" style="text-align: justify;"><a data-cke-saved-href="https://www.neb.com/tools-and-resources/usage-guidelines/cloning-guide" href="https://www.neb.com/tools-and-resources/usage-guidelines/cloning-guide">PCR-based cloning</a><br></li><li class="font_7" style="text-align: justify;">GelPilot DNA Molecular Weight Markers<br></li><li class="font_7" style="text-align: justify;">QIAquick Gel Extraction Kit<br></li></ol></li></ol> |
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− | <ol><li class="font_7" style="text-align: justify;">HT-22 cells were lysed in RIPA buffer with phosphate and protease inhibitors<br></li><li class="font_7" style="text-align: justify;">Protein concentration was measured for each OR<br></li><li class="font_7" style="text-align: justify;">Proteins were prepared with | + | <ol><li class="font_7" style="text-align: justify;">HT-22 cells were lysed in RIPA buffer with phosphate and protease inhibitors<br></li><li class="font_7" style="text-align: justify;">Protein concentration was measured for each OR<br></li><li class="font_7" style="text-align: justify;">Proteins were prepared with Laemmli sample buffer before loading into 4% Bis-Tris gels to measure protein purity and size<br></li><li class="font_7" style="text-align: justify;">Relevant documents<ol><li class="font_7" style="text-align: justify;"><a data-cke-saved-href="https://www.mcdb.ucla.edu/Research/Banerjee/protocols/maxiprep--Qiagen.pdf" href="https://www.mcdb.ucla.edu/Research/Banerjee/protocols/maxiprep--Qiagen.pdf"></a><a data-cke-saved-href="https://2017.igem.org/File:Western_Blotting_Protocol.docx" href="https://2017.igem.org/File:Western_Blotting_Protocol.docx">Western Blotting Protocol<a data-cke-saved-href="https://www.mcdb.ucla.edu/Research/Banerjee/protocols/maxiprep--Qiagen.pdf" href="https://www.mcdb.ucla.edu/Research/Banerjee/protocols/maxiprep--Qiagen.pdf"> </a><br></li></ol></li></ol> |
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− | <ol><li class="font_7" style="text-align: justify;">Nucleofected HT-22 cells were stimulated with individual odorants<br></li><li class="font_7" style="text-align: justify;">The neurons generated action potentials that were measured via a microelectrode array<br></li><li class="font_7" style="text-align: justify;">Relevant documents<ol><li class="font_7" style="text-align: justify;">Staining with Voltage Sensitive Dyes<a data-cke-saved-href="https://www.mcdb.ucla.edu/Research/Banerjee/protocols/maxiprep--Qiagen.pdf" href="https://www.mcdb.ucla.edu/Research/Banerjee/protocols/maxiprep--Qiagen.pdf"> </a><br></li></ol></li></ol> | + | <ol><li class="font_7" style="text-align: justify;">Nucleofected HT-22 cells were stimulated with individual odorants<br></li><li class="font_7" style="text-align: justify;">The neurons generated action potentials that were measured via a microelectrode array<br></li><li class="font_7" style="text-align: justify;">Relevant documents<ol><li class="font_7" style="text-align: justify;"><a data-cke-saved-href="https://2017.igem.org/File:Calcium_Dye_Fluorescence.docx" href="https://2017.igem.org/File:Calcium_Dye_Fluorescence.docx">Staining with Voltage Sensitive Dyes<a data-cke-saved-href="https://www.mcdb.ucla.edu/Research/Banerjee/protocols/maxiprep--Qiagen.pdf" href="https://www.mcdb.ucla.edu/Research/Banerjee/protocols/maxiprep--Qiagen.pdf"> </a><br></li></ol></li></ol> |
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Latest revision as of 23:15, 1 November 2017
USMA-West_Point