Primers used in RNAse protection assay
| cDNA | Size (bp) | Upstream primer (5′-3′) | Downstream primer No 1 (5′-3′) | Downstream primer No 2 (5′-3′) |
| IL-1α | 407 | CAA GGA GAG CAT GGT GGT AGT AGC AAC CAA CG | GCA CTG GTT GGT CTT CAT CTT GGG C | TAG TGC CGT GAG TTT CCC AGA AGA AGA GGA GG |
| IL-1β | 348 | GCT ACG AAT CTC CGA CCA CCA CTA CAG C | CCT TGT ACA AAG GAC ATG GAG AAC ACC | CTT ATC ATC TTT CAA CAC GCA GGA CAG G |
| IL-1 RA | 308 | CCA TTC AGA GAC GAT CTG CCG ACC | GCT TGT CCT GCT TTC TGT TCT CGC | CTG TCT GAG CGG ATG AAG GCG AAG C |
| GAPDH | 188 | GAC ATC AAG AAG GTG GTG AAG CAG GC | CCA AAT TCG TTG TCA TAC CAG GAA ATG AGC |
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RNA was reverse transcribed using downstream primer No 2, and an initial PCR was performed using the resultant cDNA as template together with upstream primer and downstream primer No 2. An aliquot of this first PCR reaction was then used as template for a second round of PCR using upstream primer and downstream primer No 1 (except for construction of the GAPDH probe which required only a single round of PCR using upstream primer and downstream primer No 2). Trinucleotide repeats (not shown) containing deoxyuracil residues were added to upstream and downstream primers to facilitate rapid cloning.
