Table 1.

PCR primers and cycling conditions

Agent	Target gene	Forward primer (5′−3′)	Reverse primer (5′−3′)	Product size (nt)	AT (°C)
FULV FULV FULV	L segment M segment S segment	CTTTCTCACCTGTTGCTACC AGGACTCTGGCAACAAGGC CATACCGTCGATTCATGGCC	GGTCTAATGCAGTATCTGGC ACGACAGTATTGCGCACCAC TTACCATCGGACATCTGCCG	312  239  277	60–55 61–56 61–56
AMSV AMSV qPCR	L protein L protein	GAACCTTCTGCTGGAAGGG TTTTCCTCCGGACAATCGGC	ATCGTCCCATGAAGTCGG GTATGAGCGTAGAAGCGACC	379 157	61–56 58
C. hepatica	18S	CCCGTCCCGAACACTGTCA	CTGACCTCTTCAAGACAGGC	309	59

AT, PCR annealing temperature; range of temperatures (xx–yy) indicates touchdown PCR (delta 0.5°C/cycle for 10 cycles).