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Table 2 Primer sequences and PCR conditions

From: Oral keratinocytes support non-replicative infection and transfer of harbored HIV-1 to permissive cells

Target

Primer

Sequences (5'-3')

PCR conditions

Integrated HIV-1

   

DNA a

   

• 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

 
  1. a Primer sequences and PCR conditions were modified from [40]
  2. b Primer sequences and PCR conditions were modified from [74]