Identification of the HFE gene by PCR.

WT and C282Y cells were analyzed for HFE and HFEgene mutations by allele-specific PCR. Primers specific for wild-typeHFE (C282 and H63) and mutant HFE (Y282 and D63) were used in PCR reactions on DNA isolated from the 2 cell lines. Lanes 1 and 5, C282 primers; lanes 2 and 6, Y282 primers; lanes 3 and 7, H63 primers; lanes 4 and 8, D63 primers. The 430-bp band (larger mol. wt. band) in all lanes represents a control PCR product. The other bands (arrows) are specific for HFE. Std (far left lane) is a 100-bp ladder standard.