HIF1α / P582S = 1744 C/T / SYBR Green / Primer Common / 5’ – CAC AGA TTT AGA CTT GGA GAT G – 3’ / 63.5°C / Eppendorf
Primer WT C / 5’ – GGA ACT GCT TTC TAA TGG TG – 3’
Primer Var T / 5’ – GGA ACT GCT TTC TAA TGA TG – 3’
A588T = 1762 G/A / SYBR Green / Primer Common / 5’ – GTA GTG GTG GCA TTA GCA GTA GG – 3’ / 57.0°C / Eppendorf
Primer WT G / 5’ – CAT TAG AAA GCA GTT CCG – 3’
Primer Var A / 5’ – CAT TAG AAA GCA GTT CCA – 3’
VEGF-A / -2578 C/A / TaqMan / Primer FWD / 5’ – CAG AGG CTA TGC CAG CTG TA – 3’ / 60.0°C / Eppendorf
Primer REV / 5’ – GGT TTC TGA CCT GGC TAT TTC CA – 3’
WT C Probe / 5’ – VIC- ACC CAG ATC GTG CCA G -MGB-NFQ – 3’
Var A Probe / 5’ – 6-FAM- CAC CCA GAT CTT GCC AG -MGB-NFQ – 3’
-1498 C/T / TaqMan / Primer FWD / 5’ – GTG TGG GTG AGT GAG TGT GT – 3’ / 60.0°C / Eppendorf
Primer REV / 5’ – GTG ACC CCT GGC CTT CTC – 3’
WT C Probe / 5’ – VIC- TCC AAC GCC CTC AAC -MGB-NFQ – 3’
Var T Probe / 5’ – 6-FAM- CTC CAA CAC CCT CAA C -MGB-NFQ – 3
-1154 G/A / TaqMan / Primer FWD / 5’ – CGG GCC AGG CTT CAC T – 3’ / 62.0°C / Applied Biosystems
Primer REV / 5’ – CCG CTA CCA GCC GAC TTT TAA – 3’
WT G Probe / 5’ – VIC- CTC AGC CCC TCC ACA C -MGB-NFQ – 3’
Var A Probe / 5’ – 6-FAM- CCT CAG CCC TTC CAC AC -MGB-NFQ – 3’
-634 G/C / TaqMan / Primer FWD / 5’ – GAG AGA AGT CGA GGA AGA GAG AGA – 3’ / 56.0°C / Applied Biosystems
Primer REV / 5’ – CCC AAA AGC AGG TCA CTC ACT T -3’
WT G Probe / 5’ – VIC- CCC TGT CCC TTT CG -MGB-NFQ – 3’
Var C Probe / 5’ – 6-FAM- CCT GTC GCT TTC G -MGB-NFQ - 3’
936 C/T / TaqMan / Primer FWD / 5’ – ACT CCG GCG GAA GCA TTC – 3’ / 60.0°C / Eppendorf
Primer REV / 5’ – AGC AAG AAA AAT AAA ATG GCG AAT CCA – 3’
WT C Probe / 5’ – VIC- CAA GAG GGA CCG TGC TG -MGB-NFQ – 3’
Var T Probe / 5’ – 6-FAM- AAG AGG GAC CAT GCT G -MGB-NFQ – 3’
eNOS / -786 T/C / TaqMan / Primer FWD / 5’ – ACC AGG GCA TCA AGC TCT TC – 3’ / 60.0°C / Eppendorf
Primer REV / 5’ – GCA GGT CAG CAG AGA GAC TAG – 3’
WT T Probe / 5’ – VIC- CAG GGT CAG CCA GCC A -MGB-NFQ – 3’
Var C Probe / 5’ – 6-FAM- CAG GGT CAG CCG GCC A -MGB-NFQ – 3’
E298D = 894 G/T / TaqMan / Primer FWD / 5’ – GGC TGG ACC CCA GGA AA – 3’ / 60.0°C / Eppendorf
Primer REV / 5’ – CAC CCA GTC AAT CCC TTT GGT – 3’
WT G Probe / 5’ – VIC- CCA GAT GAG CCC CCA -MGB-NFQ – 3’
Var T Probe / 5’ – 6-FAM- CCC AGA TGA TCC CCC A -MGB-NFQ – 3’
VEGFR-1 / -962 C/T / TaqMan / Primer FWD / 5’ – GCA TCA ATG CGG CCG AAA A – 3’ / 60.0°C / Eppendorf
Primer REV / 5’ – TTT GGG AAG ACT GCG AAC CA – 3’
WT C Probe / 5’ – VIC- CAC GGA CAC GCT CC -MGB-NFQ – 3’
Var T Probe / 5’ – 6-FAM- ACG GAC ATG CTC C -MGB-NFQ – 3’
VEGFR-2 / V297I = 889 G/A / TaqMan / Primer FWD / 5’ – CAG TCT GGG AGT GAG ATG AAG AAA T– 3’ / 60.0°C / Eppendorf
Primer REV / 5’ – GCA CAG GTG TAC AAT CCT TGG T – 3’
WT G Probe / 5’ – VIC- CTA TAG ATG GTG TAA CCC -MGB-NFQ – 3’
Var A Probe / 5’ – 6-FAM- CTA TAG ATG GTA TAA CCC -MGB-NFQ – 3’
Q472H = 1416 A/T / TaqMan / Primer FWD / 5’ – TCT AAT TAA AGA AAA ATT AAA TCA TAA TAT GCG CTG TTA TCT C – 3’ / 60.0°C / Eppendorf
Primer REV / 5’ – CAA GGG TAT GGG TTT GTC ACT GA – 3’
WT A Probe / 5’ – VIC- TTC TTA TAG CCA AGC TGT C -MGB-NFQ – 3’
Var T Probe / 5’ – 6-FAM- TTC TTA TAG CCA TGC TGT C -MGB-NFQ – 3’
NRP-1 / F561L = 1683 C/G / TaqMan / Primer FWD / 5’- GCG GAC TTT TCC AGC TCT CT – 3’ / 60.0°C / Eppendorf
Primer REV / 5’ – CCC AGT CCG CCA TGA GT – 3’
WT C Probe / 5’ – VIC-CCT GAT GAA TCG CG-MGB-NFQ – 3
Var G Probe / 5’ – 6-FAM-CCT GAT CAA TCG CG-MGB-NFQ – 3’
V733I = 2197 G/A / SYBR Green / Primer Common / 5’- CGC CTG GTG AGC CCT GT-3’ / 62.6°C / Eppendorf
Primer WT G / 5’- ACC CTG AGT GTG CCG AC -3’
Primer Var A / 5’- ACC CTG AGT GTG CCG AT -3’
NRP-2 / K123R = 368 A/G / TaqMan / Primer FWD / 5’ – TCA TCT CCT CGG GCT CCA T – 3’ / 60.0°C / Eppendorf
Primer REV / 5’ – CCT GCC GGG CGT AGT C – 3’
WT A Probe / 5’ – VIC- CTC TAC ATC AAG TTC AC -MGB-NFQ – 3’
Var G Probe / 5’ – 6-FAM- CTC TAC ATC AGG TTC AC -MGB-NFQ – 3’
Eppendorf:Eppendorf® HotMasterMix (Cat.# 954140199)
Applied Biosystems: Applied Biosystems TaqMan Universal PCR Master Mix (Cat.# 4304437)