Team:UCSF UCB/test

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<path class = "cogs" fill="none" stroke="#000000" stroke-width="3" stroke-miterlimit="10" d="M101.422 80.159l0.923-4.613l-5.189-2.998 c-3.056-1.558-0.577-7.842 3.228-5.882c0.173 0.1 5.3 3.2 5.3 3.229l3.171-3.056l-3.518-6.169 c-1.442-3.401 4.382-6.227 5.997-3.344c1.614 2.9 3.5 6.3 3.5 6.343l5.834-1.499v-6.343c0-3.689 6.735-3.574 6.678-0.634 c-0.057 2.941-0.436 7.091-0.436 7.091l4.596 1.672l3.421-5.649c1.731-2.999 8.1 0.1 6.2 3.2 c-1.959 2.998-3.931 5.998-3.931 5.998l3.34 3.573l5.533-2.997c2.825-1.558 6.3 3.9 3.3 5.8 c-3.056 1.903-5.903 3.091-5.903 3.091l0.771 4.462l6.055 0.173c4.266-0.116 4.4 7 0.5 6.8 c-3.632-0.214-6.516-0.288-6.516-0.288l-1.904 4.554l4.485 2.435c3.185 1.7 1.2 8.752-3.39 6.2 c-4.67-2.595-4.67-2.595-4.67-2.595l-3.46 2.883l2.48 5.131c1.613 3.113-3.921 6.803-5.824 3.2 c-1.901-3.633-2.421-5.65-2.421-5.65l-3.92 1.153l-0.405 6.054c-0.403 3.056-6.572 4.382-6.63-0.172 c-0.059-4.557 0.577-5.767 0.577-5.767l-5.017-1.556l-3.481 5.86c-1.478 2.673-7.589-0.555-6.148-3.149 c1.441-2.596 3.979-6.285 3.979-6.285l-2.421-2.537l-6.688 3.575c-2.768 1.613-5.593-3.979-2.594-6.054 c2.997-2.076 5.305-2.739 5.305-2.739l-1.096-5.333h-7.38c-2.941 0-3.691-6.372 0.058-6.659c3.726-0.397 7.729-0.536 7.729-0.516 H101.422z M114.139 75.495c-5.021 5.021-5.021 13.2 0 18.181c5.021 5 13.1 5 18.2 0c5.02-5.02 5.02-13.161 0-18.181 c-5.035-5.008-13.167-5.008-18.167 0H114.139z"/>
 
-
</g>
 
-
</g>
 
-
</svg>
 
-
 
-
</a></li>
 
-
                    </ul>
 
-
                </nav>
 
-
            </header>
 
-
        </div>
 
-
        <div id = "notebooks">
 
-
            <div class="notebooks-container">
 
-
                <div class = "notebooks wrapper clearfix menu">
 
-
                    <h1>Notebooks</h1>
 
-
                    <a href = "#protocols"><h2>Protocols<svg class="down"><use xlink:href="#icon-arrow-down"></use></svg></h2></a>
 
-
                </div>
 
-
            </div>
 
-
            <div class="main-container multi-container">
 
-
                <div class="wrapper clearfix">
 
-
                    <aside>
 
-
                        <h3>Lab Members</h3>
 
-
                        <ul id = "lab-members">
 
-
                            <li><a href = "#e-amidei">Eleanor Amidei</a></li>
 
-
                            <li><a href = "#s-chu">Sabrina Chu</a></li>
 
-
                            <li><a href = "#j-hsueh">Jessica Hsueh</a></li>
 
-
                            <li><a href = "#d-lee">Derrick Lee</a></li>
 
-
                            <li><a href = "#j-shu">Jeffery Shu</a></li>
 
-
                            <li><a href = "#h-shuaixin">He Shuaixin</a></li>
 
-
                            <li><a href = "#e-wong">Eric Wong</a></li>
 
-
                            <li><a href = "#r-wong">Robert Wong</a></li>
 
-
                            <li><a href = "#i-xi">Ianto Lin Xi</a></li>
 
-
                            <li><a href = "#g-yip">George Yip</a></li>
 
-
                        </ul>
 
-
                    </aside>
 
-
                    <article id = "member-select">
 
-
                        <header><svg><use xlink:href="#icon-arrow-left"></use></svg>Select a Lab Member</header>
 
-
                    </article>
 
-
                    <article id = "e-amidei">
 
-
                        <header >
 
-
                            <h2>Eleanor Amidei</h2>
 
-
                        </header>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev2</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev3</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev3</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev3</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev3</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                </article>
 
-
                    <article id = "s-chu">
 
-
                    <header>
 
-
                        <h2>Sabrina Chu</h2>
 
-
                    </header>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev2</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev3</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                </article>
 
-
                    <article id = "j-hsueh">
 
-
                    <header>
 
-
                        <h2>Jessica Hsueh</h2>
 
-
                    </header>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev2</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev3</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev2</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev3</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                </article>
 
-
                    <article id = "d-lee">
 
-
                    <header>
 
-
                        <h2>Derrick Lee</h2>
 
-
                    </header>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev2</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev3</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev2</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev3</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev2</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev3</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                </article>
 
-
                    <article id = "j-shu">
 
-
                    <header>
 
-
                        <h2>Jeffery Shu</h2>
 
-
                    </header>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev2</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev3</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                </article>
 
-
                    <article id = "h-shuaixin">
 
-
                    <header>
 
-
                        <h2>He Shuaixin</h2>
 
-
                    </header>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev2</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev3</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                </article>
 
-
                    <article id = "e-wong">
 
-
                    <header>
 
-
                        <h2>Eric Wong</h2>
 
-
                    </header>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev2</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev3</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                </article>
 
-
                    <article id = "r-wong">
 
-
                    <header>
 
-
                        <h2>Robert Wong</h2>
 
-
                    </header>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev2</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev3</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                </article>
 
-
                <article id = "l-xi">
 
-
 
-
{{:Team:UCSF_UCB/ianto}}
 
-
 
-
</article>
 
-
                    <article id = "g-yip">
 
-
                    <header>
 
-
                        <h2>George Yip</h2>
 
-
                    </header>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev2</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                    <section>
 
-
                        <h3>Lab Day whatev3</h3>
 
-
                        <p>Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aliquam sodales urna non odio egestas tempor. Nunc vel vehicula ante. Etiam bibendum iaculis libero, eget molestie nisl pharetra in. In semper consequat est, eu porta velit mollis nec. Curabitur posuere enim eget turpis feugiat tempor. Etiam ullamcorper lorem dapibus velit suscipit ultrices. Proin in est sed erat facilisis pharetra.</p>
 
-
                    </section>
 
-
                </article>
 
-
 
-
            </div> <!-- #main -->
 
-
            </div> <!-- #main-container -->
 
-
        </div>
 
-
       
 
-
        <div id = "protocols">
 
-
            <div class="notebooks-container">
 
-
                <div class = "notebooks wrapper clearfix menu">
 
-
                    <a href = "#notebooks"><h2>Notebooks<svg class="up"><use xlink:href="#icon-arrow-up"></use></svg></h2></a>
 
-
                    <h1>Protocols</h1>
 
-
                </div>
 
-
            </div>
 
-
            <div  class="main-container multi-container">
 
-
                <div class="wrapper clearfix">
 
-
                <aside>
 
-
                    <h3>Protocols</h3>
 
-
                    <ul id = "lab-members">
 
-
                        <li><a href = "#miniprep">Minipreps</a></li>
 
-
                        <li><a href = "#pcr-pure">PCR Purification</a></li>
 
-
                        <li><a href = "#g-extract">Gel Extractions</a></li>
 
-
                        <li><a href = "#e-transform">E-Coli Transformation</a></li>
 
-
                        <li><a href = "#y-transform">Yeast Transformation</a></li>
 
-
                        <li><a href = "#e-pcr">E-Coli Colony PCR</a></li>
 
-
                        <li><a href = "#y-pcr">Yeast Colony PCR</a></li>
 
-
                       
 
-
                        <li><a href = "#g-stocks">Yeast Frozen Glycerol Stocks</a></li>
 
-
                    </ul>
 
-
                </aside>
 
-
                <article id = "member-select">
 
-
                    <header>Select a Protocol<svg><use xlink:href="#icon-arrow-right"></use></svg></header>
 
-
                </article>
 
-
                <article id = "miniprep">
 
-
                    <h3>Miniprep</h3>
 
-
                    <ol>
 
-
                    <li>Resuspend pelleted bacterial cells in 250 µl Buffer PI and transfer to a microcentrifuge tube.</li>
 
-
                    <li>Add 250 µl Buffer P2 and gently invert tube 4-6 times to mix</li>
 
-
                    <li>Add 350 µl Buffer N3 and invert immediately 4-6 times</li>
 
-
                    <li>Centrifuge for 10 min at 13,000 rpm</li>
 
-
                    <li>Add supernatant to QIAprep spin column</li>
 
-
                    <li>Centrifuge for 30-60s - Discard flow through</li>
 
-
                    <li>Wash QIAprep spin column by adding 0.5 ml Buffer PB and centrifuging for 30-60s - Discard flow through</li>
 
-
                    <li>Wash column by adding 0.75 ml Buffer PE and centrifuginh for 30-60s</li>
 
-
                    <li>Discard flow through and centrifuge for an additional 1 min to remove residual wash buffer</li>
 
-
                    <li>Place column in clean 1.5 ml microcentrifuge tube. To elute DNA add 50 µl water or buffer EB, let stand for 1 min and centrifuge for 1 min.</li>
 
-
                    </ol>
 
-
                </article>
 
-
                <article id = "pcr-pure">             
 
-
                    <h3>PCR Purification</h3>
 
-
 
-
                    <ol>
 
-
                    <li>5 volumes of Buffer PB (with ethanol) to 1 volume PCR sample</li>
 
-
                    <li>Apply sample to column and spin for 1 min</li>
 
-
                    <li>Discard flow through </li>
 
-
                    <li>To wash add 0.75 ml of Buffer PE to column and centrifuge for 1 min</li>
 
-
                    <li>Discard flow through and place column back in the same tube</li>
 
-
                    <li>Cetrifuge for an addtitional minute (Dry Spin)</li>
 
-
                    <li>Place column in clean 1.5 ml tube</li>
 
-
                    <li>[Elute DNA] Add 30µl water to column, let stand for 1 min and centrifuge for 1 min </li>
 
-
                    </ol>   
 
-
                </article>
 
-
                <article id = "g-extract">
 
-
                    <h3>Gel Extraction</h3>
 
-
                    <p>QlAquick Gel Extraction Kit </p>
 
-
                    <ol>
 
-
                    <li>Cut Gel</li>
 
-
                    <li>Weigh it in a colorless tube</li>
 
-
                    <li>Add 3 volumes Buffer Q G to 1 volime Gel (100mg ~ 100µl)</li>
 
-
                    <li>Incubate @ 50° C for 10 min or until completely dissolved (vortex every 2-3 min to help dissolve)</li>
 
-
                    <li>Add 1 gel volume isopropanal to the sample and mix</li>
 
-
                    <li>Place a QlAquick soin column in a provided 2ml collection tube</li>
 
-
                    <li>Place sample in column &amp; spin for 1 min --&gt; discard flow through</li>
 
-
                    <li>To wash add 0.75 ml Buffer PE to column &amp; centrifue for 1 min, then dry spin</li>
 
-
                    <li>Place column in 1.5 ml tube</li>
 
-
                    <li>Add 35µl H2O &amp; centrifuge for 1 min.</li>
 
-
                    </ol>
 
-
                </article>
 
-
                <article id = "e-transform">                   
 
-
                    <h3>E. Coli Transformation (Digestion/Ligation)</h3>
 
-
 
-
                    <pre><code>
 
-
- 10 µl ligation
 
-
- 50 µl competent cells
 
-
--&gt; 30m on ice
 
-
--&gt; 45s heatshock 42° C
 
-
--&gt; 2m on ice
 
-
- 250 µl of SOC media
 
-
--&gt; 1h shake 37° C
 
-
                    </code></pre>   
 
-
                </article>
 
-
                <article id = "y-transform">
 
-
                   
 
-
<h3>Yeast Transformation</h3>
 
-
 
-
<table><thead>
 
-
<tr>
 
-
<th>Reagents</th>
 
-
</tr>
 
-
</thead><tbody>
 
-
<tr>
 
-
<td>YPD</td>
 
-
</tr>
 
-
<tr>
 
-
<td>1 M LiOAc</td>
 
-
</tr>
 
-
<tr>
 
-
<td>10X TE pH 7.5</td>
 
-
</tr>
 
-
<tr>
 
-
<td>1X TE pH 7.5, 0.1 M LiOAc</td>
 
-
</tr>
 
-
<tr>
 
-
<td>50% PEG 3350</td>
 
-
</tr>
 
-
<tr>
 
-
<td>DMSO</td>
 
-
</tr>
 
-
<tr>
 
-
<td>Salmon Sperm DNA (ssDNA)</td>
 
-
</tr>
 
-
</tbody></table>
 
-
 
-
<ul>
 
-
<li>PEG = viscous, pipette slow</li>
 
-
<li>boil ssDNA aliquots </li>
 
-
</ul>
 
-
 
-
<p>Previous Day : Grow yeast strain to be transformed in 5-10 mL YPD overnight at 30° C</p>
 
-
 
<ol>
<ol>
-
<li>Set up digest to linearize DNA </li>
+
<li>First</li>
-
<li>Dilute O/N culture ~ 1:20 in YPD grow 2-4 hours at 30° C</li>
+
<li>Second</li>
-
<li>Prepare ssDNA - boil for 10m cool on ice for at least 10m (10 µl of 10 mg/mL stock per transformation)</li>
+
</ol>  
-
<li>Harvest cells in centrifuge - 3000 rpm, 2-5 min</li>
+
-
<li>Wash with 1 ml 0.1 M LiOAin TE</li>
+
-
<li>Pellet cells - 3000 rpm , 2-5 min</li>
+
-
<li>Resuspend pellet in 100 µl 0.1 M LiOAc in TE per 2.5 ml culture, split into 100 µl per epindorph tube for each transformation</li>
+
-
<li>to 100 µl cells add 100 µg ssDNA (10 µl of 10 mg/mL sock), 1-5 µl target DNA</li>
+
-
<li>Add (in order): 480 µl 50% PEG 3350, 60 µl 10X TE, 60 µl 1 M LiOAc (for final 40% PEG, 1X TE, 0.1 M LiOAc)  <em>Optional</em>: Add 75 µl DMSO <em>definitely did</em></li>
+
-
<li>Vortex</li>
+
-
<li>Incubate 42° C for 30m &amp; begin drying plates</li>
+
-
<li>Pellet (6000 rpm - 2m), discard supernatant (remove PEG completely by pipetting), resuspend in 500 µl YPD (or selective media)</li>
+
-
<li>Pellet, discard supernatant, resuspend in risidual ~ 50 µl YPD</li>
+
-
<li>Plate on selective media</li>
+
-
<li>Incubate 1-3 days</li>
+
-
</ol>   
+
-
                </article>
+
-
                <article id = "e-pcr">
+
-
                    <h3>Colony PCR for screening E. Coli</h3>
+
-
 
+
-
<p>Pick single colonies ( 5 or so from each plate ) mix in 25 µl H2O in a tube. Use 5 µl in PCR reaction</p>
+
-
 
+
-
<table><thead>
+
-
<tr>
+
-
<th>Reagents</th>
+
-
<th>1X</th>
+
-
<th>6X</th>
+
-
</tr>
+
-
</thead><tbody>
+
-
<tr>
+
-
<td>2X GoTaq Green PCR Master Mix</td>
+
-
<td>10 µl</td>
+
-
<td>60 µl</td>
+
-
</tr>
+
-
<tr>
+
-
<td>10 µM Forward primer</td>
+
-
<td>1 µl</td>
+
-
<td>6 µl</td>
+
-
</tr>
+
-
<tr>
+
-
<td>10 µM Reverse primer</td>
+
-
<td>1 µl</td>
+
-
<td>6 µl</td>
+
-
</tr>
+
-
<tr>
+
-
<td>Water</td>
+
-
<td>3 µl</td>
+
-
<td>18 µl</td>
+
-
</tr>
+
-
<tr>
+
-
<td>Bacterial cells (template)</td>
+
-
<td>5 µl</td>
+
-
<td>-----</td>
+
-
</tr>
+
-
</tbody></table>
+
-
 
+
-
<p>Cycle (Varies): </p>
+
-
 
+
-
<pre><code>95° C | 5m
+
-
 
+
-
  30x:
+
-
        95° C | 45s
+
-
        55° C | 30s
+
-
        72° C | 1m per kb
+
-
 
+
-
72° C | 10m
+
-
4° C | Forever
+
-
</code></pre>
+
-
 
+
-
<p>load 5 µl onto gel</p>
+
-
 
+
-
<p>for all positive bands - take the rest of the bands and inoculate them into an overnight LB (+antibiotic) for miniprep</p>
+
-
                    </article>
+
-
                <article id = "y-pcr">
+
-
                  <h3>Colony PCR for Yeast &amp; Patching</h3>
+
-
 
+
-
                    <ol>
+
-
                    <li>Number colonies</li>
+
-
                    <li>Patch on plate</li>
+
-
                    <li>Mix in 10 µl NaOH</li>
+
-
                    <li>Boil for 20m</li>
+
-
                    <li>PCR</li>
+
-
                    </ol>
+
-
 
+
-
                    <p>Cycle (varies): </p>
+
-
                    <pre><code>
+
-
95° C | 5m
+
-
30x:
+
-
        95° C | 45s
+
-
        55° C | 30s
+
-
        72° C | 1m per kb
+
-
 
+
-
72° C | 10m
+
-
4° C | Forever
+
-
                    </code></pre>
+
-
                </article>
+
-
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+
-
                    <h3>Frozen Glycerol Stocks Yeast</h3>
+
-
 
+
-
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+
-
                    <li>Grow Overnight in YPD (2-5 mL) then dilute 1:20 in YPD, grow to OD 0.4-0.5</li>
+
-
                    <li> Add 350 µl cells to 350 µl sterile 60% glycerol in cryovial, vortex to mix, snap freeze in liquid nitrogen and store at -80° C (but actually we just stuck it in the freezer because nobody will let us use liquid nitrogen).</li>
+
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Latest revision as of 02:59, 15 October 2014

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