Table 1

Characteristics of the two multiplex MLVA assays
Locus Primers MLVA primer sequence and label (5′-3′)   Repeat sequence Repeat length (bp)   Sequencing primers (5′-3′)
Multiplex assay 1
Ms27 (Cox2) Forward primer NED-CGCTATTTTTTCAGTTTTGAGTAA TGAAGA 6 CAGTGCGTCCAGATTTCATTG
Reverse primer GTCGCAAACGTCGCACT GTCCGGATACAGCTTGAAAAGT
Ms28 (Cox5) Forward primer VIC-CGATGACGACAAAAAAGACT TAAGAA 6 GCAGAAAAAGAACATGAATGTGATTGTG
Reverse primer GCGGTAATTACTGTAAATAAATACAAAGACA GCCGGTAMCCTTCTCTAAATATTGCAA
Ms34 (Cox1) Forward primer 6FAM-ATCAGCGACTCGAAGAAAAA GAAAAG 6 CCAGTATCTCGTACGTCTCRATTT
Reverse primer AGGGTGACTTTTTCACTTAAAG CGTTTGAACACGCAACTGTTTT
Multiplex assay 2
Ms20b Forward primer VIC-TGTAACAGCACCGCCTGA GGAAGAAGCGCCACCCG 33 ACAGGTGAGTCGCCATTAACG
Reverse primer GCTTTGCCCTTTCCTTGATTTTC AAATAGGGTTGCCTCC CCATTGGGATCAAGTTCATGACTAT
Ms24 (Cox4) Forward primer NED-CCATTGTGTAATTGACATGAAGAA GACGGAA 7 TATGCGCATCTTCTCGGAGCA
Reverse primer GCCACACAACTCTGTTTTCAG GCGCTCCTTCCTCCTGTAAG
Ms31 (Cox7) Forward primer 6FAM-CCGGTATTCTAACCAACTGAAC CAGAGGA 7 AGATAAAAAGAAAAAGCAACCCGTGAA
Reverse primer GAATCCCTCAGCACCCATTC GGGTGCGTTTCCAAAAATAGTATAGG

Per assay, three different primer sets, labeled with different fluorescent dyes, target three different loci. Loci are indicated by their previously published names: Ms [9], and Cox [10]. The sequencing primer sets were used for sequencing MLVA motifs.

de Bruin et al.

de Bruin et al. BMC Veterinary Research 2012 8:165   doi:10.1186/1746-6148-8-165

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