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Sequencing

160 bytes removed, 15:46, 8 December 2007
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<p>In genetics and biochemistry, <strong>sequencing</strong> means to determine the primary structure (or primary sequence) of an unbranched biopolymer. Sequencing results in a symbolic linear depiction known as a <strong>sequence</strong> which succinctly summarizes much of the atomic-level structure of the sequenced molecule.</p>
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<h2><span class="mw-headline">DNA sequencing</span></h2>
<h3><span class="mw-headline">Sanger sequencing</span></h3>
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<div class="thumbinner" style="WIDTH: 162px"><img class="thumbimage" height="332" alt="Part of a radioactively labelled sequencing gel" width="160" border="0" src="http://upload.wikimedia.org/wikipedia/commons/c/cb/Sequencing.jpg" width="160" border="0" />
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Part of a radioactively labelled sequencing gel</div>
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<p>In chain terminator sequencing (Sanger sequencing), extension is initiated at a specific site on the template DNA by using a short oligonucleotide 'primer' complementary to the template at that region. The oligonucleotide primer is extended using a DNA polymerase, an enzyme that replicates DNA. Included with the primer and DNA polymerase are the four deoxynucleotide bases (DNA building blocks), along with a low concentration of a chain terminating nucleotide (most commonly a <strong>di-</strong>deoxynucleotide). Limited incorporation of the chain terminating nucleotide by the DNA polymerase results in a series of related DNA fragments that are terminated only at positions where that particular nucleotide is used. The fragments are then size-separated by electrophoresis in a slab polyacrylamide gel, or more commonly now, in a narrow glass tube (capillary) filled with a viscous polymer.</p>
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<div class="thumbinner" style="WIDTH: 182px"><img class="thumbimage" height="42" alt="View of the start of an example dye-terminator read (click to expand)" width="180" border="0" src="http://upload.wikimedia.org/wikipedia/commons/thumb/4/44/Sanger_sequencing_read_display.gif/180px-Sanger_sequencing_read_display.gif" width="180" border="0" />
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View of the start of an example dye-terminator read (click to expand)</div>
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<li id="_note-0"><strong><a title="" href="http://en.wikipedia.org/wiki/Sequencing#_ref-0">^</a></strong> <a class="external text" title="http://www.stenutz.eu/sop" rel="nofollow" href="http://www.stenutz.eu/sop" rel="nofollow">A practical guide to structural analysis of carbohydrates</a> </li>
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