Rolling circle amplification: A (random) primer on the enrichment of an infinite linear DNA template
Author(s) -
Kieser Rachel E.,
Budowle Bruce
Publication year - 2019
Publication title -
wiley interdisciplinary reviews: forensic science
Language(s) - English
Resource type - Journals
ISSN - 2573-9468
DOI - 10.1002/wfs2.1359
Subject(s) - dna , rolling circle replication , multiple displacement amplification , applications of pcr , primer (cosmetics) , computational biology , biology , dna sequencing , genetics , polymerase chain reaction , dna polymerase , microbiology and biotechnology , chemistry , gene , digital polymerase chain reaction , dna extraction , organic chemistry
Abstract Rolling circle amplification (RCA) is a robust enzymatic process in which a circular DNA molecule serves as a template for exponential amplification of molecular targets of interest. Random primers that have bound to DNA sequences across the circularized DNA template initiate copying via the action of a highly processive DNA polymerase. Following a cascade of strand displacement events, long tandem copies of the original template sequence are generated. Because the template molecule is in a circular format, it essentially serves as an infinite linear template. In conjunction with selective enrichment techniques, RCA may prove highly useful in the selectivity and sensitivity of current DNA typing systems. In addition, circularization potentially may allow for amplification of fragmented and low copy number DNA encountered in forensic casework samples that can be subsequently typed. This article is categorized under: Forensic Biology > Forensic DNA Technologies