Table 1.
Primers for amplification of DPOL, pTP and hexon gene sequences
| Primer-set abbreviation | Targeted gene | Name of primer | sequence 5'-3' | PCR length | Annealing temp. |
|---|---|---|---|---|---|
| Degenerate primers | |||||
| DPOL-cons | DPOL | 4431-s$ | GTnTwyGAyAThTGyGGhATGTAyGC | ||
| 4428-as | GAGGCTGTCCGTrTC(n/i)CCGTA# | 956 bp | 45°C | ||
| 4428-s | CGGACGCCTCTGyTGGAC(n/i)AA | ||||
| 4429-as | GGCCAGCACrAA(n/i)GArGC | 650 bp | 45°C | ||
| Hex-5'-cons | Hexon | 4515-s | GTGGATGG(n/i)GA(r/i)GG(n/i)TACA | ||
| 4515-as | CGCACAACGTC(r/i)AA(n/i)AC(y/i)TC | 536 bp | 45°C | ||
| 4516-s | TGTAACATGAC(y/i)AA(r(i)GA(y/i)TGG | ||||
| 4516-as | CAGGGCCCCCAT(n/i)GACA | 381 bp | 45°C | ||
| Hex3'-cons | Hexon | 4517-s | CGCAATGGTC(n/i)TACATGCAC | ||
| 4517-as | CAGTGCCCGA(r/i)TA(k/i)GG(n/i)TT | 340 bp | 45°C | ||
| 4518-s | GCAGGACGC(y/i)TCGGAGTA | ||||
| 4518-as | CACCC(k/i)GTT(r/i)TC(n/i)CC | 230 bp | 45°C | ||
| pTP-cons | pTP | 4521-s | TGGCGACGT(n/i)GT(n/i)TACAG | ||
| 4521-as | CGGACT(y/i)(k/i)GA(r/i)CCTGAAA | 260 bp | 45°C | ||
| 4522-s | TACAGCCG(n/i)GTSTGGAAC | ||||
| 4522-as | CTGAAAGAGAGTTC(n/i)ACAGAATCA | 230 bp | 45°C | ||
| Specific primers for long-distance PCR | |||||
| pTP-DPOL-LD | pTP, DPOL | 4659-s | TCGCATCTCCAACGACCT | ||
| 4659-as | GCATCCATGGTGAAGATTCC | 2350 bp | 60°C | ||
| Hex-LD | Hexon | 4618-s | AGTTCGCTACACACTGGCTG | ||
| 4618-as | ATTGCGGTGATGATTGAATG | 1354 bp | 59°C | ||
| pTP-Hex-LD | pTP, Hexon | 4662-s | CTCGGTATCGTTGACGGC | ||
| 4662-as | GATCAACGGGCACAAAGC | 10044 bp | 60°C | ||
| Primers specific for AdV species B | |||||
| Hex-loop2 | Hexon | 5442s | GAACAAGATACTTTAGCATGTGGAA | ||
| 5442as | GATTGAATGGATTAACATTGTCC | 468 bp | 55°C | ||
| Hexon | 5443s | TAGAAAATCACGGGGTGGAAGA | |||
| 5443as | GGCATCCAAAGACCATCTG | 380 bp | 55°C | ||
$ s = sense, as = antisense # I = inosine