Differential expression of RANK, RANK-L, and osteoprotegerin by synovial fluid neutrophils from patients with rheumatoid arthritis and by healthy human blood neutrophils

Table 1 DNA sequences of the forward and reverse primers for the qualitative and semi-quantitative reverse transcriptase-polymerase chain reaction analyses

Gene identity	Accession number	Primer sequences^a	Annealing temperature (°C)	Number of cycles (Quan.)^b	Number of cycles (Qual.)^c	Size of PCR product (bp)
RANK-L	AF019047	5'-CTG-ATG-AAA-GGA-GGA-AGC-AC-3'	65	29	35	546
		5'-GAT-GAC-ACC-CTC-TCC-ACT-TC-3'
OPG	U94332	5'-TGC-TGT-TCC-TAC-AAA-GTT-TAC-G-3'	56	35	40	433
		5'-CTT-TGA-GTG-CTT-TAG-TGC-GTG-3'
RANK	AF018253	5'-CCT-GGA-CCA-ACT-GTA-CCT-TC-3'	58	34	40	500
		5'-TTC-CTC-TAT-CTC-GGT-CTT-GC-3'
TRAF6	U78798	5'-TGA-TAG-TGT-GGG-TGG-AAC-TG-3'	58	27	35	456
		5'-CTC-CTT-GGA-CAA-TCC-TTC-AG-3'
β-Actin	NM001101	5'-CGT-GAC-ATT-AAG-GAG-AAG-CTG-TGC-3'	58	21	28	375
		5'-CTC-AGG-AGG-AGC-AAT-GAT-CTT-GAT-3'

^aFor each pair of sequences, the forward primer appears first and the reverse primer appears second. ^bThe number of cycles used in the semi-quantitative (quan.) PCR experiments corresponds to the linear phase of the amplification reaction. ^cThe number of cycles used in the qualitative (qual.) PCR experiments was increased to allow the possible detection of the mRNA. OPG, osteoprotegerin; PCR, polymerase chain reaction; RANK, receptor activator of nuclear factor-kappa-B; RANK-L, ligand of receptor activator of nuclear factor-kappa-B; TRAF6, tumor necrosis factor receptor-associated factor 6.

ISSN: 1478-6362