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

Table 4 Primer sequences and detailed PCR conditions used to generate standard recombinant DNA.

Gene name		5'-3' primer sequence	Position^a (cDNA)	Annealing temperature^b
IL1a	FW	GTGGTGGTGTCAGCAACATCAAAC	275–862	56°C
	RW	GAAATCTATCATGGAGGGCAGTCC
IL1b	FW	TGAAAGCTCTCCACCTCAATGGAC	501–894	57°C
	RW	TGCAGCCATCTTTAGGAAGACACG
IL1RA	FW	AAGACCTTCTACCTGAGGAACAACC	139–310	55°C
	RW	GCTTGGTGTCATCTCCAGACTTG
IL1R1	FW	TGTCTACTGGAAGTGGAATGGGTC	1143–1500	56°C
	RW	GGGAAGAAAATCAGAGCAGGAGTC
IL1R2	FW	CACCCAGTTCTTGGAGACGATTG	226–598	57°C
	RW	TGGAGGAGAGAGCTGAGATTTGC
IL6	FW	TCTGGAGTTCCGTTTCTACCTGG	388–682	55°C
	RW	CATAGCACACTAGGTTTGCCGAG
IL6R	FW	AGCAGGCAATGCTACCATTCAC	264–873	57°C
	RW	GTCGGTATCGAAGCTCGAATTG
TNFa	FW	AGCACAGAAAGCATGATCCGAG	4–499	58°C
	RW	CCTGGTATGAAGTGGCAAATCG
TNFR2	FW	TCAGATGTGCTGTGCTAAGTGTCC	93–512	58°C
	RW	GCCAGGATGCTACAAATGCG
SOCS3	FW	ATGGTCACCCACAGCAAGTTTC	18–679	56°C
	RW	TACTGGTCCAGGAACTCCCGAATG

Abbreviations: see Tables 1, 2. These primer sets allow generating recombinant DNA to ensure the specificity of the PCR amplification or to generate standard curves. ^a Position of amplification product within cDNA sequence. Genbank accession number are given in Tables 1, 2, ^b melting temperature of specific PCR product.

ISSN: 1471-2172