Random amplification of polymorphic DNA: Difference between revisions

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Revision as of 02:06, 6 October 2021

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{{Refimprove|date=June 2016}}

'''Random amplification of polymorphic DNA''' ('''RAPD'''), pronounced "rapid",<ref>{{Cite web|title=Random Amplified Polymorphic DNA (RAPD)|url=https://www.ncbi.nlm.nih.gov/probe/docs/techrapd/|access-date=2020-11-10|website=www.ncbi.nlm.nih.gov}}</ref> is a type of [[polymerase chain reaction]] (PCR), but the segments of DNA that are amplified are random.<ref>{{Cite web|title=rDNA: Random Amplification of Polymorphic DNA (RAPD)|url=http://www.rvc.ac.uk/review/dna_1/5_RAPD.cfm|access-date=2016-06-03|website=www.rvc.ac.uk}}</ref> The scientist performing RAPD creates several arbitrary, short primers (8–12 nucleotides), then proceeds with the PCR using a large template of genomic DNA, hoping that fragments will amplify. By resolving the resulting patterns, a semi-unique profile can be gleaned from an RAPD reaction.

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==How it works==

Unlike traditional PCR analysis, RAPD does not require any specific knowledge of the DNA sequence of the target organism: the identical 10-mer primers will or will not amplify a segment of DNA, depending on positions that are complementary to the primers' sequence. For example, no fragment is produced if primers annealed too far apart or 3' ends of the primers are not facing each other. Therefore, if a mutation has occurred in the template DNA at the site that was previously complementary to the primer, a PCR product will not be produced, resulting in a different pattern of amplified DNA segments on the gel.

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* PCR is an enzymatic reaction, therefore, the quality and concentration of template DNA, concentrations of PCR components, and the PCR cycling conditions may greatly influence the outcome. Thus, the RAPD technique is notoriously laboratory dependent and needs carefully developed laboratory protocols to be reproducible.

* Mismatches between the primer and the template may result in the total absence of PCR product as well as in a merely decreased amount of the product. Thus, the RAPD results can be difficult to interpret, unlike traditional PCR analysis.

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== See also ==

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==External links==

* {{MeshName|RAPD+Technique}}

{{PCR}}

[[Category:Biochemistry detection methods]]