Primers used for reverse transcription-polymerase chain reaction analysis of cyclooxygenase (COX) isoforms and β-actin mRNA expression in human and mouse colonic tissues
| Gene | Nucleotide sequence | Product size (bp) |
|---|---|---|
| After three minutes of denaturation at 94°C, PCR was performed as follows: 30 cycles of denaturation at 94°C (one minute), annealing at 50°C (1.5 minutes), extension at 72°C (two minutes), final extension at 72°C for 10 minutes (human COX-1, COX-2); 32 cycles of denaturation at 94°C (15 seconds), annealing at 60°C (15 seconds), extension at 72°C (one minute), final extension at 72°C for seven minutes (mouse COX-1); 35 cycles of denaturation at 94°C (15 seconds), annealing at 55°C (15 seconds), extension at 72°C (one minute), final extension at 72°C for seven minutes (mouse COX-2). S, sense; A, antisense. | ||
| Human COX-1 | S: 5′-TGC CCA GCT CCT GGC CCG CCG CTT-3′ | 301 |
| A: 5′-GTG CAT CAA CAC AGG CGC CTC TTC-3′ | ||
| Human COX-2 | S: 5′-TTC AAA TGA GAT TGT GGG AAA ATT GCT-3′ | 305 |
| A: 5′-AGA TCA TCT CTG CCT GAG TAT CTT-3′ | ||
| Mouse COX-1 | S: 5′-AGG AGA TGG CTG CTG AGT TGG-3′ | 602 |
| A: 5′-AAT CTG ACT TTC TGA GTT GCC-3′ | ||
| Mouse COX-2 | S: 5′-ACA CAC TCT ATC ACT GGC ACC-3′ | 274 |
| A: 5′-TTC AGG GAG AAG CGT TTG C-3′ | ||
| Human and mouse β-actin | S: 5′-TCA TGA AGT GTG ACG TTG ACA TCC GT-3′’ | 286 |
| A: 5′-CTT AGA AGC ATT TGC GGT GCA CGA TG-3′ | ||