Table 1 Wolbachia primers for all molecular assays. The assays include next-generation multilocus sequence typing (NGMLST), polymerase chain reaction (PCR), and real-time quantitative PCR (qPCR). The primers are organized according to the analysis aim and the target gene. Exact sequences and amplicon sizes are indicated. For the NGMLST, the binding regions are indicated in bold. The reaction conditions are provided in the supplementary text S3.
Aim | Gene | Forward primer | Reverse primer | Size (bp) |
|---|---|---|---|---|
NGMLST | gatB | 5’-ACACTGACGACATGGTTCTACA GAKTTAAAYCGYGCAGGBGTT-3’ | 5’-TACGGTAGCAGAGACTTG GTCTTGGYAAYTCRGGYAAA GA-3’ | 447 |
coxA | 5’-ACACTGACGACATGGTTCTACA AGTCACTATATTCAATATGCGTGCAAAAGGT-3’ | 5’-TACGGTAGCAGAGACTT GGTCTRCCAGTTATAACGCC AATAAAAATTGTTGTAG-3’ | 454 | |
hcpA | 5’-ACACTGACGACATGGTTCTACA GGCTGCCTGATCCCGAACTCA-3’ | 5’-TACGGTAGCAGAGACTTG GTCTCTCCAAATTTTGTA TAGAAAGCGTCACGTACT-3’ | 468 | |
ftsZ | 5’-ACACTGACGACATGGTTCTACA GGGTGGYGGWACYGGAACAGGTGC-3’ | 5’-TACGGTAGCAGAGACTTG GTCTTGCAGCRCTRATWGM CCTATCTTCTCC-3’ | 445 | |
fbpA | 5’-ACACTGACGACATGGTTC TACAYGGKGCTKCAACTTAT KCTGG-3’ | 5’-TACGGTAGCAGAGACTTG GTCTYACTATTCTYTTYCYT GCAAARCAAGA-3’ | 423 | |
PCR complementary to NGMLST | coxA wSc2 | 5’-ATTGTAGCTTTGCCAGTGCT-3’ | 5’-TGCATAAACCATACCC ATATAACCAAA-3’ | 240 |
hcpA wSc2 | 5’-GCTGCACGCAAGGAAAATTT-3’ | 5’-TGCCGAAATCTTTCA TTCACA − 3’ | 388 | |
Wolbachia-general qPCR | fbpA | 5′-CCTGGTTCTGCTAAGTGCTTYGATATG-3′ | 5′- GCTGTTTCACCTTCTTTGGAAATTC-3′ | 89 |
wSc1-specific qPCR | fbpA | 5’-TGGAAGAAGCTCGCGAAATT-3’ | 5’-CTTCACCGCGTGGAT AAGAC-3’ | 81 |
wSc2-specific qPCR | fbpA | 5’-CTGAGGCTAAGTCTTGCGGA-3’ | 5’-AAAGCCGCTATATGTG CAGC-3’ | 121 |
wSc1-specific qPCR | gatB | 5’-GCACATTTGGCACTCGCT-3’ | 5’-CCAAAGTGACGTCAAACAGT AAGG-3’ | 150 |
wSc2-specific qPCR | gatB | 5’-GGAAAGTGGAGGGGAGATAAGT-3’ | 5’-TCAACAGGCAATAAATCAG GTTCT-3’ | 132 |
wSc1-specific qPCR | coxA | 5’-TCGCTCACTAAGATGCCACTA-3’ | 5’-CACCAGCAAGTACCGGTA AG-3’ | 85 |
wSc2-specific qPCR | coxA | 5’-TGTAGCTTTGCCAGTGCTTG-3’ | 5’-TACAGGATCACCACCACCAG-3’ | 100 |
wSc1-specific qPCR | hcpA | 5’-AATATGAAGGCTGCGGACCT-3’ | 5’-TCCCAAATTACCGCCTTTGC-3’ | 119 |
wSc2-specific qPCR | hcpA | 5’-GTGGTGTTGCAGGAGAGAATTA-3’ | 5’-ACGCACTTCAGAAGCAGT-3’ | 119 |
wSc1-specific qPCR | ftsZ | 5’-CAAAAGCAGCCAGAGAAGCA-3’ | 5’-GCACACCTTCAAAACCGAAC-3’ | 108 |
wSc2-specific qPCR | ftsZ | 5’-TCTGCACATTGGTATCAGAGGA-3’ | 5’-TTGCCTTGCCCATTTCACTC-3’ | 107 |
