Primers (G4+RS1) were used to generate fragment 1 (124 bp) for identification of the nucleotide−28 (A–Gwhereas primers (FG3+G3) with 134 bp in fragment 2 was used for identification of codon 17
(A–T), codon 26 (G–A or Hb E), IVSI-1 (G–T) and IVSI-5(G–C) mutations.