Difference between revisions of "Team:NYU Shanghai/Protocols"
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<h4>Recipes</h4> | <h4>Recipes</h4> | ||
− | <div id="Plate" class="collapsed | + | <div id="Plate" class="collapsed"> |
− | <h5 style="display:inline-block"><span class="noselect">LB-Agar Plates</span></h5> | + | <h5 style="display:inline-block" onclick="expandPlate()"><span class="noselect">LB-Agar Plates</span></h5> |
<br> | <br> | ||
<div id="PlateText" style="display:none"> | <div id="PlateText" style="display:none"> | ||
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</div> | </div> | ||
− | <div id="LB" class="collapsed | + | <div id="LB" class="collapsed"> |
− | <h5 style="display:inline-block"><span class="noselect">LB Broth</span></h5> | + | <h5 style="display:inline-block" onclick="expandLB()"><span class="noselect">LB Broth</span></h5> |
<br> | <br> | ||
<div id="LBText" style="display:none"> | <div id="LBText" style="display:none"> | ||
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</div> | </div> | ||
− | <div id="SOC" class="collapsed | + | <div id="SOC" class="collapsed"> |
− | <h5 style="display:inline-block"><span class="noselect">SOC Media</span></h5> | + | <h5 style="display:inline-block" onclick="expandSOC()"><span class="noselect">SOC Media</span></h5> |
<br> | <br> | ||
<div id="SOCText" style="display:none"> | <div id="SOCText" style="display:none"> | ||
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<h4>Making Color</h4> | <h4>Making Color</h4> | ||
− | <div id="luc" class="collapsed | + | <div id="luc" class="collapsed"> |
− | <h5 style="display:inline-block"><span class="noselect">Luciferase</span></h5> | + | <h5 style="display:inline-block" onclick="expandluc()"><span class="noselect">Luciferase</span></h5> |
<br> | <br> | ||
<div id="lucText" style="display:none"> | <div id="lucText" style="display:none"> | ||
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<li>D-Luciferin free acid</li> | <li>D-Luciferin free acid</li> | ||
<li>ATP</li> | <li>ATP</li> | ||
− | <li>MgSO4 · 7H2O</li> | + | <li>MgSO4 · 7H2O</li> |
<li>1M HEPES Buffer</li> | <li>1M HEPES Buffer</li> | ||
<li>Lysozyme</li> | <li>Lysozyme</li> | ||
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<p>Prepare using ATP free water. Combine 1 mM luciferin or luciferin salt, 3 mM ATP and 15 mM MgSO4 in 30mM HEPES buffer, pH 7.8. Store substrate solution at -20ºC in polypropylene or glass.</p> | <p>Prepare using ATP free water. Combine 1 mM luciferin or luciferin salt, 3 mM ATP and 15 mM MgSO4 in 30mM HEPES buffer, pH 7.8. Store substrate solution at -20ºC in polypropylene or glass.</p> | ||
<b>Preparing 1mM D-Luciferin:</b> | <b>Preparing 1mM D-Luciferin:</b> | ||
− | <p>Directions for a 5 mg sample: Dissolve 0.034 gr dithiothreitol in 22 mL of QH2O. Add 0.2 mL of this DTT solution to the 5 mg d-luciferin. Add 4 uL of 10M NaOH to dissolve the luciferin. Dilute this into the remaining 21.8 mL of DTT solution and store as aliquots at - 80 ºC in darkness until use.</p> | + | <p>Directions for a 5 mg sample: Dissolve 0.034 gr dithiothreitol in 22 mL of QH2O. Add 0.2 mL of this DTT solution to the 5 mg d-luciferin. Add 4 uL of 10M NaOH to dissolve the luciferin. Dilute this into the remaining 21.8 mL of DTT solution and store as aliquots at - 80 ºC in darkness until use.</p><br> |
<b>Procedure:</b><br> | <b>Procedure:</b><br> | ||
Bacterial lysis: | Bacterial lysis: | ||
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<li>In a dark room, add about 250-350 uL of reagent solution to each sample of lysis product.</li> | <li>In a dark room, add about 250-350 uL of reagent solution to each sample of lysis product.</li> | ||
<li>Light should be emitted within two-three seconds.</li> | <li>Light should be emitted within two-three seconds.</li> | ||
− | </ol> | + | </ol><br> |
<b>Example Calculations:</b><br> | <b>Example Calculations:</b><br> | ||
Lysis Buffer (Desired Total Volume: 15mL)<br> | Lysis Buffer (Desired Total Volume: 15mL)<br> | ||
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</div> | </div> | ||
− | <div id="chromo" class="collapsed | + | <div id="chromo" class="collapsed"> |
− | <h5 style="display:inline-block"><span class="noselect">Chromoproteins</span></h5> | + | <h5 style="display:inline-block" onclick="expandchromo()"><span class="noselect">Chromoproteins</span></h5> |
<br> | <br> | ||
<div id="chromoText" style="display:none"> | <div id="chromoText" style="display:none"> | ||
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<div id="assembly" class="column"> | <div id="assembly" class="column"> | ||
<h4>3A Assembly</h4> | <h4>3A Assembly</h4> | ||
− | <div id="transform" class="collapsed | + | <div id="transform" class="collapsed"> |
− | <h5 style="display:inline-block"><span class="noselect">Transformation</span></h5> | + | <h5 style="display:inline-block" onclick="expandtransform()"><span class="noselect">Transformation</span></h5> |
<br> | <br> | ||
<div id="transformText" style="display:none"> | <div id="transformText" style="display:none"> | ||
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</div> | </div> | ||
− | <div id="mp" class="collapsed | + | <div id="mp" class="collapsed"> |
− | <h5 style="display:inline-block"><span class="noselect">Miniprep</span></h5> | + | <h5 style="display:inline-block" onclick="expandmp()"><span class="noselect">Miniprep</span></h5> |
<br> | <br> | ||
<div id="mpText" style="display:none"> | <div id="mpText" style="display:none"> | ||
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</div> | </div> | ||
− | <div id="rd" class="collapsed | + | <div id="rd" class="collapsed"> |
− | <h5 style="display:inline-block"><span class="noselect">Restriction Digest</span></h5> | + | <h5 style="display:inline-block" onclick="expandrd()"><span class="noselect">Restriction Digest</span></h5> |
<br> | <br> | ||
<div id="rdText" style="display:none"> | <div id="rdText" style="display:none"> | ||
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</div> | </div> | ||
− | <div id="ligate" class="collapsed | + | <div id="ligate" class="collapsed"> |
− | <h5 style="display:inline-block"><span class="noselect">Ligation</span></h5> | + | <h5 style="display:inline-block" onclick="expandligate()"><span class="noselect">Ligation</span></h5> |
<br> | <br> | ||
<div id="ligateText" style="display:none"> | <div id="ligateText" style="display:none"> | ||
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</div> | </div> | ||
− | <div id="gel" class="collapsed | + | <div id="gel" class="collapsed"> |
− | <h5 style="display:inline-block"><span class="noselect">Gel Electrophoresis</span></h5> | + | <h5 style="display:inline-block" onclick="expandgel()"><span class="noselect">Gel Electrophoresis</span></h5> |
<br> | <br> | ||
<div id="gelText" style="display:none"> | <div id="gelText" style="display:none"> | ||
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</div> | </div> | ||
− | <div id="gelex" class="collapsed | + | <div id="gelex" class="collapsed"> |
− | <h5 style="display:inline-block"><span class="noselect">Gel Extraction</span></h5> | + | <h5 style="display:inline-block" onclick="expandgelex()"><span class="noselect">Gel Extraction</span></h5> |
<br> | <br> | ||
<div id="gelexText" style="display:none"> | <div id="gelexText" style="display:none"> | ||
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</div> | </div> | ||
− | <div id="PCR" class="collapsed | + | <div id="PCR" class="collapsed"> |
− | <h5 style="display:inline-block"><span class="noselect">PCR</span></h5> | + | <h5 style="display:inline-block" onclick="expandPCR()"><span class="noselect">PCR</span></h5> |
<br> | <br> | ||
<div id="PCRText" style="display:none"> | <div id="PCRText" style="display:none"> | ||
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</div> | </div> | ||
− | <div id="PCRclean" class="collapsed | + | <div id="PCRclean" class="collapsed"> |
− | <h5 style="display:inline-block"><span class="noselect">PCR Cleanup</span></h5> | + | <h5 style="display:inline-block" onclick="expandPCRclean()"><span class="noselect">PCR Cleanup</span></h5> |
<br> | <br> | ||
<div id="PCRcleanText" style="display:none"> | <div id="PCRcleanText" style="display:none"> |
Revision as of 05:21, 16 September 2015