Detection of D835 mutations in the

FLT3 gene. To detect D835 mutations, we amplified exon 17 by PCR, and then digested it with the EcoRV endonuclease (A). The amplified products of wild type were digested to 2 bands (68 bp and 46 bp) by the EcoRV. When amplified products contained D835 mutations, undigested bands (114 bp) were visualized on agarose gel electrophoresis. M indicates the molecular weight marker (HaeIII digested pBR332 plasmid DNA) (B). The undigested bands were directly sequenced (C). In this sample, the first nucleotide G of D835 was substituted with T, resulting in an Asp to Tyr amino acid change (D835Y).