In order to determine the concentration of spiked TERT C228T and TERT C250T mutant alleles at which the nested PCR assays can detect, the genomic DNA of HCC cell lines HUH733 (C228T carrier) and Mahlavu (C250 carrier)33 were mixed against genomic DNA myeloid leukaemia cell line HL-60 (unknown TERT promoter mutation) to make serial dilutions of 50%, 25%, 10%, 1%, 0.5%, 0.1% and 0% of either TERT C228T or TERT C250T mutations. Here, TERT is linked to leukemia.