diff --git a/wiki/pages/experiments.html b/wiki/pages/experiments.html index 81e9c32deb84075f6108c8bceb8257588cc8b0f4..6de336aee383068ad59cbaa14884a0e686ce0736 100644 --- a/wiki/pages/experiments.html +++ b/wiki/pages/experiments.html @@ -153,16 +153,41 @@ <div class="h2"><i>C. reinhardtii </i> Transformation</div> <div class="h3">Materials and Reagents</div> <ul style="text-align:justify; font-family:AccidenzCommons; color:#185A4F; font-weight:400; font-size: min(1.5vw, 22px); font-style: normal; line-height: normal;"> - <li></li> - </ul> + <li>Yeast culture plate</li> + <li>200mM LiOAc, 1% SDS solution</li> + <li>96% ethanol</li> + <li>70% ethanol</li> + <li>H₂O</li> + </ul> + <div class="h3">Equipment</div> <ul style="text-align:justify; font-family:AccidenzCommons; color:#185A4F; font-weight:400; font-size: min(1.5vw, 22px); font-style: normal; line-height: normal;"> - <li></li> - </ul> + <li>Microcentrifuge</li> + <li>Vortex mixer</li> + <li>Water bath or heat block (70°C)</li> + <li>Micropipettes and sterile tips</li> + <li>Microcentrifuge tubes</li> + </ul> + <div class="h3">Procedure</div> <ol style="text-align:justify; font-family:AccidenzCommons; color:#185A4F; font-weight:400; font-size: min(1.5vw, 22px); font-style: normal; line-height: normal;"> - <li></li> - </ol> + <li>Pick one yeast colony from the plate and transfer the culture to a new plate or liquid medium.</li> + <li>With the same pipette tip, suspend cells in 100 μl of 200mM LiOAc, 1% SDS solution.</li> + <li>Incubate for 5 minutes at 70°C.</li> + <li>Add 300 μl of 96% ethanol, vortex.</li> + <li>Spin down DNA and cell debris at 15,000 g for 3 minutes.</li> + <li>Wash pellet with 70% ethanol.</li> + <li>Dissolve the pellet in 100 μl of H₂O or TE and spin down cell debris for 15 seconds at 15,000 g.</li> + <li>Prepare PCR reaction (example using GoTaq™ Green Master Mix by Promega):</li> + <ul style="text-align:justify; font-family:AccidenzCommons; color:#185A4F; font-weight:400; font-size: min(1.5vw, 22px); font-style: normal; line-height: normal;"> + <li>GoTaq™ Green Master Mix, 2X: 12.5μl</li> + <li>Forward primer, 10μM: 1μl</li> + <li>Reverse primer, 10μM: 1μl</li> + <li>DNA solution obtained from above: 1μl</li> + <li>Nuclease-free water to 25μl</li> + </ul> + </ol> + <div class="h2">Yeast-<i>E. coli</i> Fusion</div> <div class="h3">Materials and Reagents</div> <ul style="text-align:justify; font-family:AccidenzCommons; color:#185A4F; font-weight:400; font-size: min(1.5vw, 22px); font-style: normal; line-height: normal;">