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Created page with "<p>1<sup>st</sup> sequencing – Sanger sequencing</p> <ul> <li>Sanger sequencing by capillary electrophoresis is the gold-standard DNA sequencing technique.</li> <li>Base..."
<p>1<sup>st</sup> sequencing – Sanger sequencing</p>
<ul>
<li>Sanger sequencing by capillary electrophoresis is the gold-standard DNA sequencing technique.</li>
<li>Based on the selective incorporation of chain-terminating ddNTP by DNA polymerase during in vitro DNA replication.</li>
<li>requires a single-stranded DNA template, a DNA primer, a DNA polymerase, normal deoxynucleosidetriphosphates (dNTPs), and modified di-deoxynucleosidetriphosphates (ddNTPs)</li>
<li>resulted DNA fragments are heat denatured and separated by size using gel electrophoresis</li>
<li>DNA polymerases copy single-stranded DNA templates by adding nucleotides to a growing chain. Chain elongation occurs at the 3' end of a primer, an oligonucleotide that anneals to the template. The deoxynucleotides added to the extension product are selected by base-pair matching to the template.</li>
<li>Thermal cycling -> purification -> capillary electrophoresis</li>
</ul>
<p style="margin-left:38.0pt"><a href="http://media.invitrogen.com.edgesuite.net/ab/applications-technologies/pharma-biotherapeutics/DNA_sequencing.swf">http://media.invitrogen.com.edgesuite.net/ab/applications-technologies/pharma-biotherapeutics/DNA_sequencing.swf</a></p>
<p style="margin-left:38.0pt"><a href="https://en.wikipedia.org/wiki/Sanger_sequencing">https://en.wikipedia.org/wiki/Sanger_sequencing</a></p>
<ul>
<li>Sanger sequencing by capillary electrophoresis is the gold-standard DNA sequencing technique.</li>
<li>Based on the selective incorporation of chain-terminating ddNTP by DNA polymerase during in vitro DNA replication.</li>
<li>requires a single-stranded DNA template, a DNA primer, a DNA polymerase, normal deoxynucleosidetriphosphates (dNTPs), and modified di-deoxynucleosidetriphosphates (ddNTPs)</li>
<li>resulted DNA fragments are heat denatured and separated by size using gel electrophoresis</li>
<li>DNA polymerases copy single-stranded DNA templates by adding nucleotides to a growing chain. Chain elongation occurs at the 3' end of a primer, an oligonucleotide that anneals to the template. The deoxynucleotides added to the extension product are selected by base-pair matching to the template.</li>
<li>Thermal cycling -> purification -> capillary electrophoresis</li>
</ul>
<p style="margin-left:38.0pt"><a href="http://media.invitrogen.com.edgesuite.net/ab/applications-technologies/pharma-biotherapeutics/DNA_sequencing.swf">http://media.invitrogen.com.edgesuite.net/ab/applications-technologies/pharma-biotherapeutics/DNA_sequencing.swf</a></p>
<p style="margin-left:38.0pt"><a href="https://en.wikipedia.org/wiki/Sanger_sequencing">https://en.wikipedia.org/wiki/Sanger_sequencing</a></p>
