Figure 4

PCR and pyrosequencing of codons 167 and 200 of beta-tubulin isotype 1 from multiple species of cyathostomin. (a) PCR and genotype of F167Y for 13 species of cyathostomin. Lanes: M molecular weight marker Hyperladder II (Bioline), 1 and 2 Cylicocyclus nassatus, 3 and 4 Cyathostomum catinatum, 5 and 6 Cylicocyclus ashworthi, 7 and 8 Cylicostephanus goldi, 9 and 10 Cylicocyclus elongates, 11 and 12 Cylicocylus insigne, 13 Cylicostephanus longibursatus, 14 and 15 Cylicocyclus leptostomum, 16 and 17 Cyathostomum pateratum, 18 Cylicostephanus minutus, 19 and 20 Cylicodontophorus bicoronatus, 21 and 22 Coronocyclus coronatum, 23 and 24 Cylicocylcus calicatus, 25 no template control from primary round PCR, 26 second round PCR no template negative control. (b) Composite figure of PCR and genotype of F200Y for 10 species of cyathostomin. Lanes: M molecular weight marker Hyperladder II (Bioline), 1 and 2 Cyc nassatus, 3 and 4 Cys goldi, 5 and 6 Cya catinatum, 7 Cys longibursatus, 8 and 9 Cor coronatum, 10 Cys calicatus, 11 and 12 Cyc ashworthi, 13 Cylicodontophorus bicoronatus. 14 and 15 Cyc leptostomum, 16 Cys minutus, 17 no template negative control. (c) Semi-nested PCR of F167Y region for cyathostomin L3. Lanes: M molecular weight marker Hyperladder II (Bioline), 1-12 individual L3, 13 no template control. Genotypes are represented as follows:T/T, homozygous 'susceptible';A/T, heterozygous, A/A, homozygous 'resistant'.