The DNA sequence of the primers and probes
TNFα . | 5′-3′ sequence . |
|---|---|
| Human primers | |
| Sense primer | AGGAACAGCACAGGCCTTAGTG |
| Antisense primer | AAGACCCCTTCCAGATAGATGG |
| Human probe donor | GCCCCTCCACCCATGTGCTCC–FL |
| Human probe recipient | CACCCACCACCATCAGCCGCATC–AC |
| Mouse primers | |
| Sense primer | GGCTTTCCGAATTCACTGGAC |
| Antisense primer | CCCCGGCCTTCCAAATAAA |
TNFα . | 5′-3′ sequence . |
|---|---|
| Human primers | |
| Sense primer | AGGAACAGCACAGGCCTTAGTG |
| Antisense primer | AAGACCCCTTCCAGATAGATGG |
| Human probe donor | GCCCCTCCACCCATGTGCTCC–FL |
| Human probe recipient | CACCCACCACCATCAGCCGCATC–AC |
| Mouse primers | |
| Sense primer | GGCTTTCCGAATTCACTGGAC |
| Antisense primer | CCCCGGCCTTCCAAATAAA |
PCR template, primers, and HybProbes are single stranded. One HybProbe is labeled with the fluorescent donor dye fluorescein-sensor (FL), the other one is labeled with an acceptor dye (LCRed 640, Anchor [AC]). The donor dye is excited by blue light of 470 nm and emits green light of 530 nm. Nucleotide sequences are based on sequence of the human and mouse tumor necrosis factor gene (accession numbers M26331 and Y00467).33