Table 3.
HHV-6 primers/probe sets used in PCR reactions
Primers/probe sets |
Amplicon size |
|||
Nucleotide sequence 5′→3′ | DNA | cDNA | PCR cycle conditions | |
U67 HHV-6 | Ext-F- 5′-GCGTTTTCAGTGTGTAGTTCGGCAG-3′ | 524 | 92 °C:3 min, 92 °C:30 s, 55 °C:30 s, 72 °C:30 s (35 cycles), 72 °C:7 min, 4 °C | |
Ext-R- 5′-TGGCCGCATTCGTACAGATACGGAGG-3′ | ||||
Int-F- 5′-GCTAGAACGTATTTGCTGCAGAACG-3′ | 258 | 92 °C:3 min, 92 °C:30 s, 55 °C:30 s, 72 °C:30 s (35 cycles), 72 °C:7 min, 4 °C | ||
Int-R- 5′-ATCCGAAACAACTGTCTGACTGGCA-3′ | ||||
U12 HHV-6 | Ext-F-5′-CACTGTCATTGAGCTGTCCAA-3′ | 327 | 238 | 94 °C:5 min, 94 °C:30 s, 57 °C:30 s, 72 °C:30 s (35 cycles), 72 °C:7min, 4 °C |
Ext-R-5′-ACCACATGAGCACAAAATCG-3′ | ||||
Int-F-5′-CACTGTCATTGAGCTGTCCAA-3′ | 202 | 113 | 94 °C:5 min, 94 °C:30 s, 57 °C:30 s, 72 °C:30 s (35 cycles), 72 °C:7min, 4 °C | |
Int-R-5′-TGAGCGTGATTCCGGTAACT-3′ | ||||
U90 HHV-6A | F-5′-CATGAAGATGATGACAATAAAATG-3′ | 278 | 94 °C:3 min, 94 °C:20 s, 65 °C:50 s (45 cycles), 4 °C | |
R-5′-TGGAACCATCTTGTTCTGTCC-3′ | ||||
FAM-CCGCCCAGATCTGTCACTGAGGCG-TAMRA | ||||
U90 HHV-6B | F-5′-GAGACCGGGTCTGGACAACA-3′ | 145 | 94 °C:3 min, 94 °C:20 s, 65 °C:50 s (45 cycles), 4 °C | |
R-5′-GAGTTGCTGAGTTGGTAAAGG-3′ | ||||
FAM-CTCCAAGTGTACCGAAACGCTTCCTGG-TAMRA | ||||
β-Actin | F-5′-CACACTGTGCCCATCTACGA-3′ | 107 | 94 °C:3 min, 94 °C:20 s, 65 °C:50 s (45 cycles), 4 °C | |
R-5′-CTCAGTGAGGATCTTCATGAGGTAGT-3′ | ||||
FAM-ATGCCCTCCCCCATGCCATCCTGCGT-TAMRA |