Quantitative PCR and RT PCR Primers and Probes

Quantitative PCR and RT PCR Primers and Probes

<p>Supplementary Table 1. Primer and probe sequences</p><p>(Reverse Transcriptase) Quantitative PCR primers and probes GFP (Q-PCR) GFP-TQ-F CAA CAG CCA CAA CGT CTA TAT CAT GFP-TQ-R ATG TTG TGG CGG ATC TTG AAG GFP-TQ-P 5′-FAM-CCG ACA AGC AGA AGA ACG GCA TCA A-3′TAMRA HIV-1 U5/ Gag leader (Q-PCR) HIV1-TQ-F TGT GTG CCC GTC TGT TGT GT HIV1-TQ-R GAG TCC TGC GTC GAG AGA GC HIV1-TQ-P 5’-FAM-CAG TGG CGC CCG AAC AGG GA-3’TAMRA 18S (Q-PCR and RT-Q-PCR) 18S-TQ-F TCG AGG CCC TGT AAT TGG AA 18S-TQ-R CCC TCC AAT GGA TCC TCG TT 18S-TQ-P 5’-FAM-AGT CCA CTT TAA ATC CTT-3’TAMRA IL-3 (RT-Q-PCR) IL-3-TQ-F AAC TTA ACT GTT GCC TGC CTA CAT C IL-3-TQ-R AAG TCA TCC AGA TCT CGA ATG AAG AC IL-3-TQ-P 5’-FAM-C GAA TGA CTC TGC GCT GCC AGG G-3’TAMRA Primers used to generate 18S and IL-3 inserts for plasmid DNA standards 18S-F TAC CTG GTT GAT CCT GCC AGT A 18S-R TTA CGA CTT TTA CTT CCT CTA GAT AG IL-3-F ATG GTT CTT GCC AGC TCT ACC IL-3-R TAG ATA AAT TGA TAA GAC ATT TGA TGG C</p><p>Inverse PCR primer sequences and cycling conditions Lentiviral inverse PCR (1st round) 95°C 15’; 35x (94°C 45”, 53°C 30”, 72°C 1’); 72°C 10’ HUE AGC CAA TGA AGG AGA GAA CAC CC HUB GAT CAA GGA TAT CTT GTC TTC GT Lentiviral inverse PCR (Nested) 95°C 15’; 30x (94°C 45”, 54°C 30”, 72°C 1’); 72°C 10’ HUC CTT GTT ACA CCC TGT GAG CCT IP3 TCT TGT CTT CGT TGG GAG TGA Gammaretroviral inverse PCR (1st round) 95°C 15’; 35x (94°C 45”, 51°C 30”, 72°C 1’); 72°C 10’ MspI-5F GAG AAG TTC AGA TCA AGG TC MspI-5R TCC ATG CCT TGC AAA ATG GC Gammaretroviral inverse PCR (Nested) 95°C 15’; 30x (94°C 45”, 53°C 30”, 72°C 1’); 72°C 10’ Nested-5F CAA GGT CAG GAA CAG ATG GA Nested-5R GCT TGC CAA ACC TAC AGG TG Linker mediated PCR primer sequences and cycling conditions LM PCR Linker Plus NlaIII Plus GTA ATA CGA CTC ACT ATA GGG CTC CGC TTA AGG GAC TAC ATG TaqI/MspI Plus GTA ATA CGA CTC ACT ATA GGG CTC CGC TTA AGG GAC LM PCR Linker Minus NlaIII Minus PO4-TA GTC CCT TAA GCG GAG TaqI/MspI Minus PO4-GC GTC CCT TAA GCG GAG CSGW LM PCR (1st round) 95°C 5’; 30x (94°C 45”, 55°C 30”, 72°C 1’); 72°C 10’ Linker 1 GTA ATA CGA CTC ACT ATA GGG C HIV-1 3’LTR 1 AGT GCT TCA AGT AGT GTG TGC C CSGW LM PCR (Nested) 95°C 5’; 30x (94°C 45”, 55°C 30”, 72°C 1’); 72°C 10’ Linker Nested AGG GCT CCG CTT AAG GGA C HIV-1 3’LTR GTC TGT TGT GTG ACT CTG GTA AC Nested</p><p>Reverse transcriptase PCR and 5’ rapid amplification of cDNA ends and Northern blot probe primer sequences GHR RT PCR GHR A1 TGG ACA GAA GGA GAT AAT CCT GAT GHR RT3 GAA CTC GCT GTA CTT TTC AAA GCT C 5’RACE UAP CTACTACTACTAGGCCACGCGTCGACTAGTA 5’RACE - R AATAGATCCTGGGGTCTTTAAATCAG Northern Blot Ghr exon 11-3’UTR probe GHR B1 CAGCTGTTACGACCCTGACATT GHR BP1 GAATCCATATCCACTCTCATGCTC</p><p>Multiplex PCR primer sequences Name Locus-specific forward primer (Used in combination with HIV-1 U3 primer IP3 TCT TGT CTT CGT TGG GAG TGA) 108-HV-F TGC TCA GGT TGT CTC AGA GTA 110-HV-F GGA GGC AAA GCC TCT TGA TAA 112-HV-F GCT AAG CTT GGA TTT ATG CGC T 114-HV-F GGC CAA CGT TTG ATG GAG TAA T 116-HV-F GGT AGA CTT TAG AGG TAC TGA G 118-HV-F GTT CAT GCT AAG GTG AGT GAA G 120-HV-F AGC CTT AGT ATA CAG TGG GCT A</p>

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