Table 3 Oligonucleotide primers and PCR conditions for the characterization of subolesin and pathogen-specific gene expression.
From: Subolesin expression in response to pathogen infection in ticks
Gene descriptiona | Upstream/downstream primer sequences (5'-3') | PCR annealing conditions |
|---|---|---|
D. variabilis subolesin [9] | CCAGCCTCTGTTCACCTTTC CCGCTTCTGAATTTGGTCAT | 54°C, 30 sec |
R. microplus subolesin [9] | CACAGTCCGAGTGGCAGAT GATGCACTGGTGACGAGAGA | 55°C, 30 sec |
A. marginale msp4[29] | GGGAGCTCCTATGAATTACAGAGAATTGTTTAC CCGGATCCTTAGCTGAACAGGAATCTTGC | 60°C, 1 min |
A. phagocytophilum msp4[9] | GACGTGCTGCACACAGATTT CTCATCAAATAGCCCGTGGT | 54°C, 1 min |
E. canis 16S (M73221) | GTGGCAGACGGGTGAGTAAT GCTGATCGTCCTCTCAGACC | 57°C, 30 sec |
B. subtilis dal[35] | AATTGAAAGGGACCGACATC- TTAATGGTTTCGAGCCTTCC | 59°C, 30 sec |
E. coli dxs[36] | CGAGAAACTGGCGATCCTTA CTTCATCAAGCGGTTTCACA | 60°C, 30 sec |
P. pastoris CTA 1 (AB472085) | CCTGAAGGACGCCAATATGT GCTTTCCAGCCTCTTCATTG | 57°C, 30 sec |
Tick 16S rRNA [9] | GACAAGAAGACCCTA ATCCAACATCGAGGT | 42°C, 30 sec |