TABLE 1.

Forward and reverse sets of primers used for mRNA amplification of the cytokines or chemokinesa

PrimerNucleotide sequenceSize (bp)Tm (°C)No. of cycles
Aldolase Ab
    Forward 15′-GTG CTG GCT GCT GTC TAC-3′1845329
    Reverse 15′-AGG TGA TCC CAG TGA CAG-3′
IL-1β
    Forward5′-GTG GCA ATG AGG ATG ACT T-3′5345329
    Reverse5′-TGG GCT TAT CAT CTT TCA A-3′
IL-1ra
    Forward5′-TCC GCA GTC ACC TAA TCA-3′3705329
    Reverse5′-CTG TCT GAG CGG ATG AAG-3′
MCP-1
    Forward5′-CAA ACT GAA GCT CGC ACT-3′3175329
    Reverse5′-GTT TGG GTT TGC TTG TCC-3′
MIP-1β
    Forward5′-GAA GCT CTG CGT GAC TGT-3′2465329
    Reverse5′-TGG ACC CAG GAT TCA CTG-3′
Aldolase Ac
    Forward 25′-GTG CTG GCT GCT GTC TAC AA-3′21856.829
    Reverse 25′-GAC GCC TCC TCC TCA CTC TG-3′
TNF-α
    Forward5′-CTT GTT CCT CAG CCT CTT CT-3′57856.829
    Reverse5′-ACT CGG CAA AGT CGA GAT AG-3′
  • a The size of the expected PCR products, the annealing temperature (melting temperature [Tm]) for each set of primers, and the number of PCR cycles used are indicated.

  • b A pair of aldolase A (internal control) primers was used for the coamplification of aldolase A, IL-1β, IL-1ra, MCP-1, and MIP-1β.

  • c A different pair of aldolase A primers was designed for the coamplification of aldolase A and TNF-α due to GC-rich sequence of TNF-α and Tm incompatibility with the rest of the cytokine gene sequences.