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Table 2 Primer sequences and annealing temperatures of Sanger sequencing reactions

From: Prevalence of tick-borne pathogens in questing Ixodes ricinus ticks in urban and suburban areas of Switzerland

Species

Primer sequences (5'–3')

Amplicon length (bp)

Amplicon position (reference sequence)

Annealing T (°C)

Reference

Fist-step PCR

    

Borrelia spp.

locus: 5S-23S intergenic spacer (forward: GAG TTC GCG GGA GAG TAG GTT ATT; reverse: TCA GGG TAC TTA GAT GGT TCA CTT CC

420

3063–3483 (JX564636.1)

64

[111]

Babesia spp.

locus: 18S ribosomal RNA gene (forward: GTC TTG TAA TTG GAA TGA TGG; reverse: TAG TTT ATG GTT AGG ACT ACG)

489

466–955 (AJ439713)

58

[112]

Rickettsia spp.

locus: 23S–5S intergenic spacer (forward: GAT AGG TCR GRT GTG GAA GCA C; reverse: TCG GGA YGG GAT CGT GTG TTT C)

388

1–388 (AY125012)

68

[113]

 Ticks

locus: cytochrome c oxidase (forward: ACW AAY CAY AAA GAC ATT GGA AC; reverse: WGG ATG CCC RAA RAA TCA AAA T)

704

1242–1946 (KF197132)

48

[114]

Second-step PCR

    

Borrelia spp.

locus: 23S-5S intergenic spacer (forward: GGA GAG TAG GTT ATT GCC AG; reverse: GGT TCA CTT CCC CTG GTA TC)

396

3073–3468 (JX56436.1)

64

Babesia spp.

locus: 18S ribosomal RNA gene (forward: GTA ATT GGA ATG ATG GTG AC; reverse: GTT AGG ACT ACG ACG GAA TC)

475

471–946 (AJ439713)

58

Rickettsia spp.

locus: 23S-5S intergenic spacer (forward: CAG TAA TGT GTG TAG CTA AC; reverse: ATC GTG TGT TTC ACT CAT GC)

356

22–378 (AY125012)

58

 Ticks

locus: cytochrome c oxidase (forward: AYC AYA AAG ACA TTG GAA CWA T; reverse: GCC CRA ARA ATC AAA ATA RAT G)

686

1245–1933 (KF197132)

48