added protocol 2

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;">