Table 1 Primers used in this work.

From: Identification of reference genes for real-time PCR cytokine gene expression studies in sheep experimentally infected with Fasciola hepatica

Targeta

Accesion numbera

Primers sequencesb 5′–3′

Primers Tm (°C)c

Amplified product length (bp)d

Amplification efficiency

References

(R2)e

TNF-α

NM_001024860.1

F 5′-CCACGCTCTTCTGCCTGCTGCACTTCGG-3′

73.1

146

0.996

Pacheco et al.33

R 5′-AACGTGGGCTACCGGCTTGTTATTTGAGGC-3′

73.6

(99.56)

IL-1β

NM_001009465.2

F 5′-GAAGCTGAGGAGCCGTGCCTACGAACA-3′

68.8

185

1.001

Pacheco et al.33

R 5′-CCAGCACCAGGGATTTTTGCTCTCTGTCC-3′

69.0

(99.85)

FoxP3

NM_001144947.1

F 5′-GCCCATCTGGCTGGGAAGATGGCCCAAACC-3′

76.5

166

0.991

Pacheco et al.33

R 5′- AGAGGTGCCTCCGCACGGCAAACAGG-3′

76.2

(99.16)

IL-10

NM_001009327.1

F 5′-TCAGCCGTGCTCTGTTGCCTGGTCTTCC-3′

73.8

124

0.999

Pacheco et al.33

R 5′- GGACGTCCCGCAGCATGTGGGGCAG-3′

73.5

(99.9)

TGF-β

NM_001009400.1

F 5′- GGGCTTTCGCCTCAGTGCCCACTGTTC-3′

73.8

151

1.009

Pacheco et al.33

R 5′- CAGAGGGGTGGCCATGAGGAGCAGG-3′

73.8

(99.9)

PGK1

NM_001142516.1

F 5′-GTGAAGGGGAAGCGGGTCGTCATGAGAG-3′

72.9

99

1.002

Pacheco et al.10

R 5′-GCTTGGAACAGCAGCCTTGATCCTCTGG-3′

72.1

(99.43)

B2M

NM_001009284.2

F 5′-CAAGACACCCGCCAGAAGATGGAAAGC-3′

71.6

180

0.989

Pacheco et al.10

R 5′-GGAGTGAACTCAGCGTGGGACAGAAGG-3′

70.3

(99.91)

RPLP0

XM_004017413.2

F 5′-CGGCTGCTGCCCGTGCTGGTGCCAT-3′

77.7

191

1.002

Pacheco et al.10

R 5′-TTCGCTGGCGCCCACCTTGTCTCCGGTC-3′

77.1

(99.81)

G6PD

NM_001093780.1

F 5′-CGGGCGAGAGCAACGAAGCACAGAGAGC-3′

74.0

161

1.004

Pacheco et al.10

R 5′-CCAGGTCCCCCGATGCACCCATGATG-3′

74.2

(99.82)

SDHA

XM_012097183.1

F 5′-CCATGAGTTTGATGCCGTGGTGGTCGGTGC-3′

70.7

184

1.006

Pacheco et al.10

R 5′-CCGCCAGTTGTCCTCCTCCATGTTCCCCA −3′

70.4

(99.51)

ACTB

NM_001009784.1

F 5′-GCGTGATGGTGGGCATGGGCCAGAAGG-3′

76.0

170

1.003

This work

R 5′-GGGGGCCACACGCAGCTCGTTGTAGAAGG-3′

75.7

(99.99)

HSP90AA1

XM_004017995.3

F 5′-GCCGCCCCTGGAAGGAGACGACGACACG-3′

71.1

130

1.005

This work

R 5′-GCCAGGCGAGCCTCGGCAGCGCTCA-3′

72.0

(99.53)

GYPC

XM_004004772.3

F 5′-TGCCCCGCACCGGCCAGCGATGAG-3′

79.2

110

1.006

This work

R 5′-GGTCGGGCTCTGGGGTCCAAAGGGGCGTC-3′

79.3

(99.85)

GUSB

XM_015103904.1

F 5′-CTTCCGCGCCGACTTCTCCGACAAC-3′

72.4

185

1.004

This work

R 5′-GCAGCGTGATCTCCCGTTCATACCACACC-3′

72.8

(99.89)

TUBB

XM_012101347.2

F 5′-CCAGGACGCCACGGCCGAGGAGGAGGG-3′

79.1

112

0.997

This work

R 5′-TGGCCGCAGCCGCGTCCCACCCCTTC-3′

79.8

(99.58)

  1. aGene symbols and accession numbers are according to the NCBI Gene database, bSequence of forward (F) and reverse (R) primers, cMelting temperature (°C) calculated by the Oligo 7 software, dPCR product size in base pair (bp), eThe real-time PCR efficiencies (E) were calculated from each efficiency curve according to the equation E = 10(−1/slope) −1. E is in the range from 0 (minimum value) to 1 (maximum and optimum). i.e. E = 1 is equal to 100% efficiency.
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