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Ribotyping
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<h2 id="technique">Technique</h2> <p>Ribotyping involves the digestion of bacterial genomic DNA with specific <a href="/facts/Restriction_enzyme/Mg0VgkCX">restriction enzymes</a>. Each restriction enzyme cuts DNA at a specific nucleotide sequence, resulting in fragments of different lengths.<a class="footnote-ref" id="fnref:4" href="#fn:4"><sup>4</sup></a> </p><p>Those fragments are then run on a <a href="/facts/Gel_electrophoresis/9J3GrsFk">Gel electrophoresis</a>, where they are separated according to size: the application of electrical field to the gel in which they are suspended causes the movement of DNA fragments (all negatively charged due to the presence of phosphate groups) through a matrix towards the positively charged end of the field. Small fragments move more easily and rapidly through the matrix, reaching a bigger distance from the starting position than larger fragments. </p><p>Following the separation in the gel matrix, the DNA fragments are moved onto nylon membranes and hybridized with a labelled 16S or 23S rRNA probe. This way only the fragments coding for such rRNA are visualised and can be analyzed.<a class="footnote-ref" id="fnref:5" href="#fn:5"><sup>5</sup></a> The pattern is then digitized and used to identify the origin of the DNA by a comparison with reference organisms in a computer database.<a class="footnote-ref" id="fnref:6" href="#fn:6"><sup>6</sup></a> </p><p>Conceptually, ribotyping is similar to probing restriction fragments of chromosomal DNA with <a href="/facts/Hybridization_probe/j3pu1L7E">cloned probes</a> (randomly cloned probes or probes derived from a specific coding sequence such as that of a <a href="/facts/Virulence_factor/jw099pff">virulence factor</a>).<a class="footnote-ref" id="fnref:7" href="#fn:7"><sup>7</sup></a> </p> <h2 id="see-also">See also</h2> <ul><li><a href="/facts/Genotyping/GRyXkRcn">Genotyping</a></li></ul> <h2 id="references">References</h2> <ol> <li id="fn:1"><p>Madigan, Michael T. (2012). Biology of Microorganisms. Pearson. p. 491. ISBN 978-0-321-73551-5. <a href="978-0-321-73551-5" target="_blank">978-0-321-73551-5</a> <a href="#fnref:1" class="footnote-back-ref">↩</a></p></li> <li id="fn:2"><p>Madigan, Michael T. (2012). Biology of Microorganisms. Pearson. p. 491. ISBN 978-0-321-73551-5. <a href="978-0-321-73551-5" target="_blank">978-0-321-73551-5</a> <a href="#fnref:2" class="footnote-back-ref">↩</a></p></li> <li id="fn:3"><p>"Investigating Microbial Diversity: Then and Now". learn.genetics.utah.edu. Retrieved 2025-04-09. <a href="https://learn.genetics.utah.edu/content/gsl/diversity/" target="_blank">https://learn.genetics.utah.edu/content/gsl/diversity/</a> <a href="#fnref:3" class="footnote-back-ref">↩</a></p></li> <li id="fn:4"><p>"Restriction Enzymes | ASU - Ask A Biologist". askabiologist.asu.edu. Retrieved 2016-03-13. <a href="https://askabiologist.asu.edu/restriction-enzymes" target="_blank">https://askabiologist.asu.edu/restriction-enzymes</a> <a href="#fnref:4" class="footnote-back-ref">↩</a></p></li> <li id="fn:5"><p>"ribotype". courses.cit.cornell.edu. Retrieved 2016-03-13. <a href="https://courses.cit.cornell.edu/biomi290/microscopycases/methods/ribotype.htm" target="_blank">https://courses.cit.cornell.edu/biomi290/microscopycases/methods/ribotype.htm</a> <a href="#fnref:5" class="footnote-back-ref">↩</a></p></li> <li id="fn:6"><p>Madigan, Michael T. (2012). Biology of Microorganisms. Pearson. p. 491. ISBN 978-0-321-73551-5. <a href="978-0-321-73551-5" target="_blank">978-0-321-73551-5</a> <a href="#fnref:6" class="footnote-back-ref">↩</a></p></li> <li id="fn:7"><p>Grimont, F., and P. A. Grimont. 1986. Ribosomal ribonucleic acid gene restriction patterns as potential taxonomic tools. Ann. Inst. Pasteur Microbiol. 137B:165–175 <a href="#fnref:7" class="footnote-back-ref">↩</a></p></li> </ol>