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Primers and Conditions
HCV 5¡¯NCR
Outer Sense
(939) CTGTGAGGAACTACTGTCTT
Outer
Anti-Sense(209) ATACTCGAGGTGCACGGTCTACGAGACCT
Inner Sense
(940) TTCACGCAGAAAGCGTCTAG
Inner
Anti-Sense (211) CACTCTCGAGCACCCTATCAGGCAGT
HGV 5¡¯NCR
Primers
Outer Sense
(4571) AGGTGGTGGATGGGTGAT
Outer
Anti-Sense (4574) TGCCACCCGCCCTCACCCGAA
Inner Sense
(4572) TGGTAGGTCGTAAATCCCGGT
Inner
Anti-Sense (4573) GGRGCTGGGTGGCCYCATGCWT
HCV 1b
ISDR primers
Outer
Sense CTCCCATGYGAGCCCGAACCGGA
Outer
Anti-Sense GGSCGYGCCCATAYGGG
Inner
Sense TCCATGCTYACCGACCC
Inner
Anti-Sense TCYTGGRTTTYCGCAGGATCTC
HIV p17
gag primers
Outer Sense
(332) GCGAGAGCGTCAGTATTAAGCGG
Outer
Anti-Sense (308) TCTGATAATGCTGAAAACATGGG
Inner Sense
(306) GGGAAAAAATTCGGTTAAGGCC
Inner
Anti-Sense (307) CTTCTACTACTTTTACCCATGC
HIV V3
primers
Outer
Sense TACAATGTACACATGGAATT
Outer
Anti-Sense ATTACAGTAGAAAAATTCCCC
Inner
Sense TGGCAGTCTAGCAGAAGAAG
Inner
Ant-Sense CTGGGTCCCCTCCTGAGG
HIV RT
Outer
Sense AGGACCTACACCTGTCAACAT
Outer
Anti-Sense ACTCAGGAATCCAGGTGGCTT
Inner
Sense GTTGACTCAGATTGGTTGCACTTT
Inner
Ant-Sense TGGACTTCATAACCCATCCAAAG
TTV
primers (hemi-nested)
Outer Sense
(30) CAGACAGAGGAGAAGGCAACATG
Inner Sense
(61) GGMAAYATGYTRTGGATAGACTGG
Anti-sense
(8230) CTGGCATTTTACCRTTTCCAAARTT
¡¡
Cycling Conditions
HCV
5¡¯NCR/HCV ISDR/HIV V3
25
cycles 94oC for 18s, 50oC for 21s, 72oC
for 90s
1
cycle 72oC at 6 min
1
cycle 20oC holding temp
HGV/HIV
p17/TTV
30
cycles 94oC for 18s, 55oC for 21s, 72oC
for 90s
1
cycle 72oC at 6 min
1
cycle 20oC holding temp
HIV RT
1st
PCR
1
cycle 94oC for 1 min
35
cycles 94oC for 1 min, 50oC for 1 min,
72oC for 1 min
1
cycle 72oC at 7 min
1
cycle 20oC holding temp
2nd
PCR
1
cycle 94oC for 1 min
30
cycles 94oC for 1 min, 53oC for 1 min,
72oC for 1 min
1
cycle 72oC at 7 min
1
cycle 20oC holding temp
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