5.08:_DNA_Sequencing_by_the_Dideoxy__Method

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<figure><img class="internal" alt="Diagram showing the conversion of ATP to methylthioadenosine. ATP loses pyrophosphate to form DMAPP, which then becomes MTA. Chemical structures and bonds are depicted throughout." loading="lazy" src="https://bio.libretexts.org/@api/deki/files/6530/ddTTP.gif?revision=1" /><figcaption>Figure 5.8.2: Sample DNA Sequencing</figcaption></figure> <p class="lt-bio-4747" style="background-color: unset;">The DNA to be sequenced is prepared as a single strand. This template DNA is supplied with</p> <ul> <li class="lt-bio-4747" style="background-color: unset;">a mixture of all four <strong>normal</strong> (deoxy) nucleotides in ample quantities <ul> <li class="lt-bio-4747" style="background-color: unset;">dATP</li> <li class="lt-bio-4747" style="background-color: unset;">dGTP</li> <li class="lt-bio-4747" style="background-color: unset;">dCTP</li> <li class="lt-bio-4747" style="background-color: unset;">dTTP</li> </ul> </li> <li class="lt-bio-4747" style="background-color: unset;">a mixture of all four <strong>dideoxy</strong>nucleotides, each present in limiting quantities and each labeled with a &quot;tag&quot; that fluoresces a different color: <ul> <li class="lt-bio-4747" style="background-color: unset;"><span style="color:green;"><strong>ddATP</strong></span></li> <li class="lt-bio-4747" style="background-color: unset;"><span class="mt-color-ff9900"><strong>ddGTP</strong></span></li> <li class="lt-bio-4747" style="background-color: unset;"><span style="color:blue;"><strong>ddCTP</strong></span></li> <li class="lt-bio-4747" style="background-color: unset;"><span style="color:red;"><strong>ddTTP</strong></span></li> </ul> </li> <li class="lt-bio-4747" style="background-color: unset;">DNA polymerase I</li>
</ul> <figure><img style="width: 620px; height: 494px;" class="internal" alt="Colorful DNA sequencing chromatogram with four rows of peaks in red, yellow, black, and gray, each representing different nucleotide bases." loading="lazy" width="620px" height="494px" src="https://bio.libretexts.org/@api/deki/files/6532/DNA_sequence.gif?revision=1&amp;size=bestfit&amp;width=620&amp;height=494" /><figcaption>Figure 5.8.3 example of a DNA sequence (455 nucleotides of the lysU gene of E. coli) courtesy of Pharmacia Biotech Inc., Piscataway, NJ</figcaption></figure>

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<hr class="autoattribution-divider" /><div class="autoattribution"><p>This page titled <a target="_blank" class="internal mt-self-link" href="/Sandboxes/johnnyphung/biology/05:_DNA/5.08:_DNA_Sequencing_by_the_Dideoxy__Method">5.8: DNA Sequencing by the Dideoxy  Method</a> is shared under a <a rel="nofollow" href="https://creativecommons.org/licenses/by/3.0" target="_blank">CC BY  3.0</a>  license and was authored, remixed, and/or curated by <a rel="nofollow" target="_blank" href="http://www.biology-pages.info/">John W. Kimball</a> via <a rel="nofollow" href="https://www.biology-pages.info/" target="_blank">source content</a> that was edited to the style and standards of the LibreTexts platform.</p></div>

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