Dilution series for mutant allele quantification
A set of mutant allele dilutions was prepared by mixing quantification cycle (Cq) normalised HEL DNA (MUT) which harbors the mutant allele only, and HL-60 DNA (WT), which harbors the wild-type allele only. In earlier experiments, the HEL DNA (MUT) had been normalised for their JAK2 copy number, based on their amplification, and mixed with genomic DNA extracted from a healthy blood donor (WT). Each MUT/WT mix was made to a final DNA concentration of 16.5 ng/μl, which corresponds to a total number of 5,000 JAK2 copies/μl.