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Table 1 Details for PCR protocols used

From: Eurasian golden jackal as host of canine vector-borne protists

Parasite

Gene

Fragment length (bp)

Primer

Primer sequence (5’-3’)

PCR conditions, master mix used

Source

Hepatozoon spp.

18S

~670

Hep F

ATACATGAGCAAAATCTCAAC

94 °C for 3 min, 35× (94 °C for 1 min, 61 °C for 1 min, 72 °C for 2 min), 72 °C for 7 min

[17]

   

Hep R

CTTATTATTCCATGCTGCAG

PPP Master Mix (TopBio s.r.o., Czech Republic)

 
 

18S

~1765

HAM 1 F

GCCAGTAGTCATATGCTTGTC

95 °C for 5 min, 35× (95 °C for 20 s, 56 °C for 1 min, 72 °C for 1 min), 72 °C for 5 min

[18]

   

HPF 2R

GACTTCTCCTTCGTCTAAG

PCRBIO Taq DNA Polymerase (PCR Biosystems Ltd, UK)

 

Piroplasmida

18S

~685

BTH 1 F

CCTGAGAAACGGCTACCA CATCT

95 °C for 10 min, 40× (95 °C for 30 s, 60 °C for 1 min, 72 °C for 1 min), 72 °C for 10 min

[19]

   

BTH 1R

TTGCGACCATACTCCCCCCA

  
  

~560

GF2

GTCTTGTAATTGGAATGA TGG

95 °C for 10 min, 40× (95 °C for 30 s, 62 °C for 1 min, 72 °C for 1 min), 72 °C for 10 min

 
   

GR2

CCAAAGACTTTGATTTCTCTC

PPP Master Mix (TopBio s.r.o., Czech Republic)

 

18S

~1730

BT1 F

GGTTGATCCTGCCAGTAGT

94 °C for 30 s, 20× (94 °C for 30 s, 65-55 °C (-0.5 °C/cycle) for 30 s, 68 °C for 1 min), 20× (94 °C for 30 s, 55 °C for 30 s, 68 °C for 1 min), 68 °C for 5 min

[20]

   

BT outer R

GGAAACCTTGTTACGACTTCTC

  
  

~1670

Piro0F2

GCCAGTAGTCATATGCTTGTCTTA

94 °C for 30 s, 20× (94 °C for 30 s, 65-55 °C (-0.5 °C/cycle) for 30 s, 68 °C for 1 min), 20× (94 °C for 30 s, 55 °C for 30 s, 68 °C for 1 min), 68 °C for 5 min

[21]

   

BT Inner R

TTC TCC TTC CTT TAA GTG ATA AG

OneTaq 2x Master Mix with standard buffer (NEB Inc., USA)

[20]

Babesia spp.

cox1

~1250

Bab_For1

ATWGGATTYTATATGAGTAT

95 °C for 1 min, 35× (95 °C for 15 s, 45 °C for 30 s, 72 °C for 30 s), 72 °C for 10 min

[22]

   

Bab_Rev1

ATAATCWGGWATYCTCCTTGG

 

modified

  

~975

Bab_For2

TCTCTWCATGGWTTAATTATGATAT

95 °C for 1 min, 35× (95 °C for 15 s, 45 °C for 30 s, 72 °C for 30 s), 72 °C for 10 min

 
   

Bab_Rev2

TAGCTCCAATTGAHARWACAAAGTG

PCRBIO Taq DNA Polymerase (PCR Biosystems Ltd, UK)

 

Theileria spp.

cox1

~1020

Cox1F133

GGAGAGCTAGGTAGTAGTGGAGATAGG

95 °C for 1 min, 35× (95 °C for 15 s, 63 °C for 15 s, 72 °C for 30 s), 72 °C for 10 min

[21]

   

Cox1R1130

GTGGAAGTGAGCTACCACATACGCTG

  
  

~985

Cox_cladeI_Fw2

GTAGTGGAGATAGGTTCATAGC

95 °C for 1 min, 35× (95 °C for 15 s, 55 °C for 15 s, 72 °C for 30 s), 72 °C for 10 min

This study

   

Cox_cladeI_Rev2

TGTATCGTGTAGTGACACGTC

PCRBIO Taq DNA Polymerase (PCR Biosystems Ltd, UK)

 

Leishmania spp.

ITS1

~280

ITS-219 F

AGCTGGATCATTTTCCGATG

qPCR: 95 °C for 5 min, 45× (95 °C for 5 s, 57 °C for 15 s, 72 °C for 15 s), melting from 60 to 95 °C at 1 °C/s

[26]

   

ITS-219R

ATCGCGACACGTTATGTGAG

iQSYBER Green Supermix (BioRad Inc., USA)

 
 

ITS1-5.8S

~300

LITSR

CTGGATCATTTTCCGATG

95 °C for 3 min, 40× (95 °C for 20 s, 53 °C for 35 s, 72 °C for 60 s), 72 °C for 5 min

[27]

   

L5.8S

ACACTCAGGTCTGTAAAC

  
     

EmeraldAmp (TaKaRa Clontech, Japan)