Abstract: Rolling circle amplification (RCA) is a method of isothermal amplification of circular DNA molecules. RCA assay involves DNA amplification using a DNA ɸ polymerase with strand displacement activity to extend a single or multiple primers annealed to a circular DNA template. The strand displacement activity allows the newly synthesized DNA template to displace the previously generated DNA molecule releasing single-stranded DNA (ssDNA). This enzymatic process of primer extension combined with DNA strand displacement generates a long single-stranded DNA containing a repeated sequence complementary to the circular DNA template. The method is successfully used for the detection of many circular single- and double-stranded DNA viruses (begomovirus, badnavirus) infecting different crops. In this chapter, different steps involved in the RCA of a begomovirus virus genome are discussed.
Publication Year: 2020
Publication Date: 2020-01-01
Language: en
Type: book-chapter
Indexed In: ['crossref']
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Cited By Count: 13
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