PROTOCOL

WAVE UPDATED WORKFLOW FOR PCR TESTING OF SURVEY SAMPLES FOR CASSAVA MOSAIC BEGOMOVIRUSES

Based on results from previous years, WAVE has adopted the following PCR primers (Table 1) and workflow (Figure 1) for the screening of all samples collected during field surveys for Cassava mosaic begomoviruses (CMB).

Table 1: Updated WAVE primers for PCR detection of Cassava mosaic begomoviruses

	Primer name	Primer sequences (5′ to 3′)	Target region	Size	Reference
1	JSP 001	ATGTCGAAGCGACCAGGAGAT	ACMV DNA-A (CP)	783 bp	Pita et al., 2001
	JSP 002	TGTTTATTAATTGCCAATACT
2	ACMVBF	TCGGGAGTGATACATGCGAAGGC	ACMV DNA-B (BV1/BC1)	628bp	Matic et al., 2012
	ACMVBR	GGCTACACCAGCTACCTGAAGCT
3	JSP 001	ATGTCGAAGCGACCAGGAGAT	EACMV DNA-A (CP)	780bp	Pita et al., 2001
	JSP 003	CCTTTATTAATTTGTCACTGC
4	CMBRepF	CRT CAA TGA CGT TGT ACC A	EACMV DNA-A (AC1)	650 bp	Alabi et al., 2008
	EACMVRepR	GGT TTG CAG AGA ACT ACA TC
5	VNF031/F	GGATACAGATAGGGTTCCCAC	EACMV-CM DNA-A (AC2/AC3)	≈ 560bp	Fondong et al., 2000
	VNF032/R	GACGAGGACAAGAATTCCAAT
6	ACMV21for	GCAGTGATGAGTTCCCCGGTGCG	DNA-A (AC3- AC2-AC1) of ACMV, EACMV, EACMCV, EACMKV, EACMMV, EACMZV and SACMV	552 bp	Matic et al., 2012
	ACMV21rev	ATTCCGCTGCGCGGCCATGGAGACC

Figure 1 : Workflow for PCR screening of WAVE survey samples for Cassava mosaic begomoviruses