Table 2. Sequences of Primers and Probes Utilized for Quantitative Real-Time PCR.
qRT-PCR was performed utilizing primers and probes specific for rhesus macaque genes. GI# - GenBank accession number; Forward – forward primer; Reverse – reverse primer; FAM – 6’ carboxyfluorescein; TAMRA - 6-carboxytetramethylrhodamine
| Gene | GI # | Real-time PCR Sequence | |
|---|---|---|---|
| Vdelta 1 (Vδ1) TCR |
339381 | Forward | 5'-TCG CCT TAA CCA TTT TAG CC-3' |
| Reverse | 5'-ACC GGA TGG TTT GGT ATG AGG T-3' | ||
| Probe | 5'-FAM-TAC AGC TAG AAG ACT CAG CAA CAT ACT TCT GTG CTC T-TAMRA-3' |
||
| Vdelta 2 (Vδ2) TCR |
37314 | Forward | 5'-GAG AAC CAG GCT GTA CTT AAG ATC CTT-3' |
| Reverse | 5'-TGA CGA AAA CGG ATG GTT TG-3' | ||
| Probe | 5'-FAM-AGA GAG AGA TGA AGG GTC TTA CTA CTG TGC CAG TG- TAMRA-3' |
||
| CD4 | 393417 | Forward | 5'-CCA CTG GAA AAA ACT CCA ACC A-3' |
| Reverse | 5'-CGA TCG CTC AGC TTG GAT G-3' | ||
| Probe | 5'-FAM-AAA GAT TCT GGG AAT TCA GGG CTC CTT CTT AAC TAA -TAMRA-3' |
||
| CCL21 / 6CKine |
74136270 | Forward | 5'-TAC CGG AAG CAG GAA CCA AG-3' |
| Reverse | 5'-CTG CTC CAT CCC AGC TAT CCT GTT CTT G-3' | ||
| Probe | SYBR Green | ||
| CCR7 | 74136330 | Forward | 5'-GGG GAA ACC AAT GAA AAG T-3' |
| Reverse | 5-GTG ACC TCA TCT TGA CAC AGG C-3' | ||
| Probe | Probe # 77 (Roche – Universal Probe Library) | ||
| CCL5 / RANTES |
74136260 | Forward | 5'-ACC AGT GGC AAG TGC TCC A-3' |
| Reverse | 5'-GGT TGG CAC ACA CTT GGC G-3' | ||
| Probe | 5'-FAM-CAA GCA GTC GTC GTC TTT GTC ACC CGA AA-TAMRA-3' | ||
| CCR5 | 116812906 | Forward | 5'-TAC CTG CTC AAC CTG GCC AT-3' |
| Reverse | 5'-TTC CAA AGT CCC ACT GGG C-3' | ||
| Probe | 5'-FAM-CCT GCT TTT CCT TCT TAC TGT CCC CTT CTG-TAMRA-3' | ||
| GAPDH | 93004459 | Forward | 5'-GCA CCA CCA ACT GCT TAG CAC-3' |
| Reverse | 5'-TCT TCT GGG TGG CAG TGA TG-3' | ||
| Probe | 5'-FAM-TCG TGG AAG GAC TCA TGA CCA CAG TCC-TAMRA-3' | ||