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Table 3 Detailed conditions used for real-time PCR assays.

From: Rat pro-inflammatory cytokine and cytokine related mRNA quantification by real-time polymerase chain reaction using SYBR green

Gene name [primer]a (μM) [Mg++]b (mM) Annealing conditionsc (°C…s) Product TMd (°C) Amplicon Lengthe (bp)
βactin 0.3 3.5 58°C…4s 83.5 97
CypA 0.4 4 58°C…5s 85.2 127
GAPDH 0.4 3 58°C…4s 86.9 202
HPRT 0.4 4 60°C…5s 84.7 123
IL1a 0.4 4 65°C…5s 81.5 85
IL1b 0.4 3.5 58°C…5s 82.0 79
IL1RA 0.4 3 65°C…5s 80.7 117
IL1R1 0.4 3 64°C…5s 80.0 105
IL1R2 0.4 4 64°C…5s 84.5 116
IL6 0.4 3 65°C…5s 80.9 79
IL6R 0.4 5 60°C…5s 84.3 117
TNFa 0.4 3 58°C…4s 84.7 111
TNFR2 0.4 4 60°C…5s 84.9* 83
SOCS3 0.4 4 65°C…5s 84.7 99
C/EBPβ 0.4 4 59°C…5s 90.2 90.2
cFos 0.4 4 60°C…5s 84.6 84.6
cJun 0.4 4 62°C…5s 89.2 89.2
COX-2 0.4 4 60°C…4s 83.8 83.8
NOS-2 0.4 4 58°C…5s 83.8 83.8
  1. Abbreviations: see Tables 1, 2.a final concentration of oligonucleotide primers (μM), b final concentration of Mg2+ (mM), c Annealing temperature (°C) and duration (s), d melting temperature of specific PCR product, e Amplicon length in base pairs. *TNF-r2: a second melting peak close to 88°C can coexist depending on template. The two peaks correspond to a single amplification product as controlled using high-resolution gel electrophoresis and PCR with recombinant and genomic DNA as templates.