Glutathione Transferase P1 Polymorphism Might Be a Risk Determinant in Heart Failure

Table 2

PCR-RFLP: primer sequences, PCR conditions, restriction enzymes, and fragment lengths.

Polymorphism	Primer sequences	PCR protocol	Gel electrophoresis results
GSTA1C-69T	F, 5-GCATCAGCTTGCCCTTCA -3; R, 5-AAACGCTGTCACCGTCCTG-3	Denature: 94°C for 3 min; Followed by 94°C for 30 s; Annealing: 56°C for 30 s; Extension: 72°C for 30 s; #cycles: 30; Final extension: 72°C for 10 min	Eam1104I incubation at 37°C overnight; GSTA1CC: 400 bp; GSTA1CT: 400 bp, 308 bp, and 92 bp; GSTA1TT: 308 bp and 92 bp
GSTP1Ile105Val	F, 5-ACCCCAGGGCTCTATGGGAA-3; R, 5-TGAGGGCACAAGAAGCCCCT-3	Denature: 95°C for 10 min; Followed by 94°C for 30 s; Annealing: 59°C for 30 s; Extension: 72°C for 30 s; #cycles: 29; Final extension: 72°C for 10 min	Alw26I incubation at 37°C overnight; GSTP1Ile/Ile: 176 bp; GSTP1Ile/Val: 176 bp, 91 bp, and 85 bp; GSTP1Val/Val: 91 bp and 85 bp
GSTM1	F, 5-GAACTCCCTGAAAAGCTAAAGC-3; R, 5-GTTGGGCTCAAATATACGGTGG-3	Multiplex PCR: Denature: 94°C for 3 min; Followed by 94°C for 30 s; Annealing: 59°C for 30 s; Extension: 72°C for 45 s; #cycles: 30; Final extension: 72°C for 4 min	GSTM1active: 215 bp band; GSTM1null: absent band
GSTT1	F, 5-TTCCTTACTGGTCCTCACATCTC-3; R, 5-TCACGGGATCATGGCCAGCA-3		GSTT1active: 480 bp band; GSTT1null: absent band
CYP1A1	F, 5-GAACTGCCACTT CAGCTGTCT-3; R, 5-CAGCTGCATTTG GAAGTGCTC-3		Successful PCR reaction: 312 bp band; Unsuccessful PCR reaction: absent band