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Using the TripleMaster PCR System for Robust Amplification of ,,, GC-Rich DNA Templates

(RB) 180 bp 75% 5' CAG GAC AGC GGC CCG GAG 3'
5 CTG CAG ACG CTC CGC CGT 3' Human apolipoprotein E gene (apo E) 268 bp 75% 5' GGC ACG GCT GTC CAA GGA 3'
5' CTC GCG GAT GGC GCT GAG 3' Human melanocortin-1 receptor gene (MC1R) 760 bp 63% 5' CTG GTG AGC TTG GTG GAG A 3'
5' GGC TGT GGA AGG CGT AGA T 3' Table 2: Final reagent concentrations in 50 ml reaction volumes. Eppendorf TripleMaster PCR System Human RB gene(180 bp) Human apo E gene (268 bp) Human MC1R gene (760 bp) Tuning Buffer with Mg2+ 1.6x; 4.0 mM 1.6x; 4.0 mM 1x; 2.5 mM DMSO 5% 5% 2% dNTP's 200 M each 200 M each 200 M each Primers 0.4 M each 0.4 M each 0.4 M each TripleMaster enzyme mix 2.0 U 2.0 U 2.0 U Template DNA 100 ng 100 ng 100 ng Invitrogen ThermalAce DNA Polymerase Kit Human RB gene(180 bp) Human apo E gene (268 bp) Human MC1R gene (760 bp) ThermalAce Buffer with Mg2+
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