Combining single-nanopore estimates of HGD65 target concentration to gauge accuracy and precision. The combined estimate data plotted at each known target concentration is the average and standard deviation of at least two estimates generated from separate single-nanopore experiments (data in Table 1). The concentrations are above the nanopore after dilution. Single-pore estimates were combined where a common scaffold/fusion reagent was used (7 pores P7-P13, and 2 pores P14-P15) or by the presence of 1% saliva background above the pore (6 pores P21-P26). The R2 value assesses how closely the average estimates are to the zero error line, with the three-parameter model (R2 = 0.985, Supplementary Table S45) outperforming the two-parameter model (R2 = 0.953, Supplementary Table S46 and Fig. S24).