Changes
From Biolecture.org
no edit summary
<p>Template preparation is to generate and amplify the template DNA, that is the main target of sequencing. there are several different ways in preparing the template DNA as much as diverse secuencing methods exist.</p>
<p> </p>
<p> </p>
<p> </p>
<p> </p>
<h2>emPCR</h2>
<p> </p>
<p> </p>
<p><img alt="" src="/ckfinder/userfiles/images/emPCR%2C%20Solid-phase%20amplification%20(2).png" style="height:183px; width:731px" /></p>
<p><strong><em> <Summary of the Em PCR></em></strong></p>
<p> </p>
<p> </p>
<p>There are primer, template DNA, dNTPs, DNA polymerase mixed with emulsion oil and water. </p>
<p> </p>
<p><img alt="" src="/ckfinder/userfiles/images/emulsion%20(2).jpg" style="height:139px; width:404px" /></p>
<p>After shaking to make emulsion, the PCR mix enters Into the emulsion(the oil dorp) and denaturation begins to seperate double strands.</p>
<p> </p>
<p> </p>
<p><img alt="" src="/ckfinder/userfiles/images/2(2).jpg" style="height:389px; width:550px" /></p>
<p>Let's call the fragment attached to the bead with adaptod(colored blue) 'reverse strand' for conveneince.</p>
<p>When the reverse strand annealed to the bead, polymerase amplifies the forward strand from the bead so that it produce one 'forward strand'.</p>
<p> </p>
<p> </p>
<p><img alt="" src="/ckfinder/userfiles/images/3(2).jpg" style="height:376px; width:550px" /></p>
<p> </p>
<p>Then the 'reverse strand', which was annealed to the bead, denatured and annealed again to the other spot.</p>
<p>And the primer also anneals to the forward strand, which was produced before.</p>
<p><img alt="" src="/ckfinder/userfiles/images/4(2).jpg" style="height:405px; width:550px" /></p>
<p>These processes so far, repeat for 30-60 cycles and generate and amplify the template DNA.</p>
<p> </p>
<p> </p>
<p> </p>
<h3>Quetions</h3>
<p>Why is the oil required?</p>
<p>Why is the sphere-shape bead required?</p>
<p> </p>
<p> </p>
<h1> </h1>
<h1> </h1>
<h1>Sequencing and imaging</h1>
<h1>Reference</h1>
<p> </p>
