Identification of mutations in G6PD gene is performed as an epidemiologic investigation of G6PD deficiency in many countries. In order to understand the hereditary background of G6PD deficiency in a population, screening of mutations is required not only in exonic regions but also for intron and promoter regions. One hundred male neonatal samples diagnosed as with G6PD deficiency by newborn screening in Singapore were used in this study. The multiplex PCR using the multiple tandem forward primers and common reverse primer (MPTP) method was carried out to detect the common 11 mutations in south-east Asia such as Gaohe 95A>G, Orissa 131C>G, Vanua-Lava 383T>C, Mahidol 487G>A, Mediterranean 563C>T, Coimbra 592C>T, Viangchan 871G>A, Chatham 1003G>A, Union 1360C>T, Canton 1376G>T and Kaiping 1388G>A. Samples whose mutations were unidentified by MPTP method were scanned at cording region, intron and promoter region by direct sequencing.Out of 100 samples, 90 samples (90.0%) were identified with one of the above mentioned common mutations. Eight out of 10 samples whose mutations were unidentified by MPTP method carried exonic mutations which had been previously reported such as Murcia 209A>G, Quing Yuan 392G>T, Nankang 517T>C, Chinese5 1024C>T. Two novel mutations were identified in these samples: one had a novel mutation (25C>T); the remaining sample carried a 49 bp deletion in intron 12.