Primer or probe name | Target | Concentration | Sequence 5′→3′ | Amplicon |
---|---|---|---|---|
Multiplex qPCR | ||||
Primer for | 23S rDNA | 0.4 μM | TTCGATGGAATGACACAGATTGT | 128 bp |
Primer rev | 23S rDNA | 0.4 μM | CCGAAAGCCCAGTCACTATC | |
Probe_hyo | B.hyodysenteriae | 100 nM | 6-FAM-CCTTAACCTTAAAGAAGCAAGCAT(BHQ-1)TTGACTCACCTCAAG-SpacerC3 | |
Probe _pilo | B.pilosicoli | 100 nM | Yakima Yellow-AGGTGATGGTTATCCTCGTCGAAT-BHQ-1 | |
Probe_ apathogen | B. intermedia/ B. innocens/ B. murdochii | 25 nM | Dragonfly Orange-CCTCAACCTTAAAGCAACAAGCAT(BHQ-2)TTTACTCATCACAAG-SpacerC3 | |
eGFP for | enhanced GFP | 0.2 μM | GACCACTACCAGCAGAACAC | 177 bp |
eGFP rev | enhanced GFP | 0.2 μM | GAACTCCAGCAGGACCATG | |
Probe_eGFP | enhanced GFP | 25 nM | ATTO 647N-AGCACCCAGTCCGCCCTGAGCA-BHQ-3 | |
Duplex PCR [12] | ||||
B.hyo_for | nox | 0.5 μM | ACTAAAGATCCTGATGTATTTG | 345 bp |
B.hyo_rev | nox | 0.5 μM | CTAATAAACGTCTGCTGC | |
B.pilo_for | 16S rDNA | 0.17 μM | AGAGGAAAGTTTTTTCGCTTC | 823 bp |
B.pilo_rev | 16S rDNA | 0.17 μM | GCACCTATGTTAAACGTCCTTG |