Random amplification of polymorphic DNA: Difference between revisions

Revision as of 15:05, 17 December 2021

Revision as of 04:10, 3 February 2022

Line 11:

==How it works==

After amplification with PCR, samples are loaded into a gel (either agarose or polyacrylamide) for [[Gel electrophoresis of nucleic acids|gel electrophoresis]]. The differing sizes created through random amplification will separate along the gel in a repeatable manner depending on the sample source. This creates a distinct DNA fingerprint.

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.