Target | Primer | Sequences (5'-3') | PCR conditions |
---|---|---|---|
Integrated HIV-1 | Â | Â | Â |
DNA a | Â | Â | Â |
• First round PCR | L-M667 | ATGCCACGTAAGCGAAACTCTGGCTAACTAGGGAACCCACTG | 95°C, 8 min and 95°C, 10 s, 60°C, 10 s, 72°C, 170 s for 12 cycles |
 | Alu 1 | TCCCAGCTACTGGGGAGGCTGAGG |  |
 | Alu 2 | GCCTCCCAAAGTGCTGGGATTACAG |  |
• Second round PCR | Lambda T | ATGCCACGTAAGCGAAACT | 95°C, 8 min and 95°C, 10 s, 60°C, 10 s, 72°C, 9 s for 40 cycles |
 | AA55M | GCTAGAGATTTTCCACACTGACTAA |  |
Linear HIV DNA a | MH531 | TGTGTGCCCGTCTGTTGTGT | 95°C, 8 min and 95°C, 10 s, 60°C, 10 s, 72°C, 6 s for 40 cycles |
 | MH532 | GAGTCCTGCGTCGAGAGAGC |  |
2-LTR circle a | HIV F | GTGCCCGTCTGTTGTGTGTGACT | 95°C, 8 min and 95°C, 10 s, 60°C, 10 s, 72°C, 10 s for 40 cycles |
 | HIV R | ACTGGTACTAGCTTGTAGCACCATCCA |  |
U5-U3 RNA a | HIV F | GTGCCCGTCTGTTGTGTGTGACT | 95°C, 2 min and 95°C, 5 s, 60°C, 10 s, 72°C, 10 s for 40 cycles |
 | HIV R | ACTGGTACTAGCTTGTAGCACCATCCA |  |
Gag | For | CCCATAGTGCAGAACATCCA | 50°C, 2 min, 95°C, 2 min, and 95°C, 15s and 60°C, 30s, for 50 cycles |
 | Rev | GGGCTGAAAGCCTTCTCTTC |  |
Singly spliced b | M669 | GTGTGCCCGTCTGTTGTGTGACTCTGGTAAC | 50°C, 2 min, 95°C, 2 min, and 95°C, 15s and 60°C, 30s, for 50 cycles |
 | La 23 | GCCTATTCTGCTATGTCGACACC |  |
Multiply spliced HIV RNA a | P659 | GACTCATCAAGTTTCTCTATCAAA | 95°C, 4 min and 95°C, 5 s, 54°C, 10 s, 72°C, 8 s for 40 cycles |
 | P413MOD | AGTCTCTCAAGCGGTGGT |  |
Unspliced HIV RNA a | La 9 | GACGCTCTCGCACCCATCTC | 95°C, 2 min and 95°C, 10 s, 60°C, 40 s for 40 cycles |
 | La 8.1 | CTGAAGCGCGCACGGCAA |  |
β-actin | Actin F | ATGGCCACGGCTGCTTCCAGC | 95°C, 15 s, 55°C, 30 s, 72°C, 15 s for 30 cycles |
 | Actin R | CATGGTGGTGCCGCCAGACAG |  |
GAPDH | GAPDH F | GAGTCAACGGATTTGGTCGT | 95°C, 15 s, 60°C, 30 s, 72°C, 15 s for 30 cycles |
 | GAPDH R | TTGATTTTGGAGGGATCTCG |  |