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Table 1 Sequences of primers used for cytokine real-time PCR (qPCR) and standard curve data

From: Immune response profile of caruncular and trophoblast cell lines infected by high- (Nc-Spain7) and low-virulence (Nc-Spain1H) isolates of Neospora caninum

Targeta

Primer

Primer sequence (5′–3′)

Product size (bp)

R 2 b

Slopec

IFN-γ (NM_174086.1)

QIFN-UPg

GATTCAAATTCCGGTGGATG

110

0.994

(−3.47)–(−3.30)

QIFN-RPg

TTCTCTTCCGCTTTCTGAGG

TNF-α (EU276079.1)

QTNF-UPg

CCAGAGGGAAGAGCAGTCC

126

0.998

(−3.39)–(−3.27)

QTNF-RPg

GGAGAGTTGATGTCGGCTAC

IL-4 (M77120.1)

QIL4-UPg

CTGCCCCAAAGAACACAACT

169

0.995

(−3.33)–(−3.54)

QIL4-RPg

GTGCTCGTCTTGGCTTCATT

IL-6 (X68723.1)

QIL-6-UPd

CTGGGTTCAATCAGGCGATT

150

0.999

(−3.22)–(−3.20)

QIL-6-RPd

GGATCTGGATCAGTGTTCTGA

IL-8 (BC103310.1)

qIL8-Fwh

CCACACCTTTCCACCCCAAA

177

0.995

(−3.36)–(−3.23)

qIL8-Rwh

CTTGCTTCTCAGCTCTCTTC

IL-10 (NM_174088.1)

QIL10-UPg

TGCTGGATGACTTTAAGGGTTACC

60

0.999

(−3.27)–(−3.42)

QIL10-RPg

AAAACTGGATCATTTCCGACAAG

IL-12p40 (NM_174356.1)

QIL12-UPg

AGTACACAGTGGAGTGTCAG

157

0.992

(−3.39)–(−3.35)

QIL12-RPg

TTCTTGGGTGGGTCTGGTTT

IL-17 (NM_001008412.1)

qIL17bov-uph

GAACTTCATCTATGTCACTGC

83

0.997

(−3.30)–(−3.18)

qIL17bov-revh

TGGACTCTGTGGGATGATGA

TGF-β1 (NM_001009400.1)

QTGF-UPd

GGTGGAATACGGCAACAAAA

117

0.999

(−3.60)–(−3.53)

QTGF-RPd

CGAGAGAGCAACACAGGTTC

TLR-2 (NM_001048231.1)

QTLR2-UPe

ACGACGCCTTTGTGTCCTAC

192

0.993

(−3.74)–(−3.38)

QTLR2-RPe

CCGAAAGCACAAAGATGGTT

ICAM-1 (NM_174348.2)

qICAM-Fwh

AGACCTATGTCCTGCCATCG

219

0.994

(−3.34)–(−3.30)

qICAM-Rwh

GGTGCCCTCCTCATTTTCCT

VCAM (XM_005204079.2)

qVCAM-Fwh

GAACTGGAAGTCTACATCTC

128

0.998

(−3.36)–(−3.32)

qVCAM-Rwh

CAGAGAATCCGTGGAGCTGG

GAPDH (NM_001034034)

GAPDH-Ff

ATCTCGCTCCTGGAAGATG

227

0.996

(−3.67)–(−3.58)

GAPDH-Rf

TCGGAGTGAACGGATTCG

β-Actin (NM_173979.3)

BACTIN-UPg

ACACCGCAACCAGTTCGCCAT

216

0.994

(−3.45)–(−3.36)

BACT216-RPg

GTCAGGATGCCTCTCTTGCT

  1. aNCBI accession numbers are for cDNA sequences used in primer design. Primer annealing was also checked with the Bos taurus genomic DNA sequences (http://www.ncbi.nlm.nih.gov/nuccore)
  2. bMinimum coefficient of regression (R2) of standard curves for each PCR target in all batches of amplification
  3. cStandard curve slopes. Minimum and maximum values for slopes for each PCR target in all batches of amplification
  4. dPrimer first described by Arraz-Solís et al. [43]
  5. ePrimer first described by Menzies & Ingham [61]
  6. fPrimer first described by Puech et al. [62]
  7. gPrimer first described by Regidor-Cerrillo et al. [9]
  8. hPrimer described in the present work for the first time