Empirical evaluation of humpback whale telomere length estimates; quality control and factors causing variability in the singleplex and multiplex qPCR methods

Table 5 Summary of quality control steps and overall assay performance for the telomere (T), reference (S), and telomere length estimates (T/S)

Criteria	Target	Assay I	Assay II	Assay III	Assay IV	Figure/Table
Serial dilution series
LDR	T	Borderline	Within range	Within range	Within range	Figure 2
	S	Borderline	Within range	Within range	Within range
NTC	T	OK	Borderline	Borderline	OK	Figures 3, 4
	S	OK	Overlap*	Overlap	Overlap*
Magnitude E	T	Intermediate	Intermediate	Intermediate	High	Figure 1
	S	Intermediate	Low	Intermediate	Intermediate
Interbatch E	T	Constant	Variable	Constant	Constant	Figure 1
	S	Constant	Variable	Constant	Constant
Dilution E	T	Variable E	No change	No change	No change	Table 3
	S	Variable E	No change	No change	No change
Precision	T	Intermediate	High	High	High	Figure 5, Table 4
	S	Intermediate	Low	Moderate	High
Accuracy	T	High	High	High	High	Table 4
	S	High	Intermediate	High	Intermediate
Resolution	T	Low	High	High	High	Table 5
	S	Intermediate	Intermediate	High	High
Precision	T/S	Intermediate	Low	Intermediate	Intermediate	Additional file 3: Figure S2
Accuracy	T/S	Low	Low	High	High	Additional file 3: Figure S2
"Unknown" samples
Outliers	T/S	Few	Many	Many	Few	Additional file 1: Table S1
Resolution	T/S	Intermediate	Low	Low	High	Additional file 1: Table S1
Quantitative method	T/S	High	Low	Intermediate	High	Additional file 5: Table S2
Inter-assay correlation	T/S	Low	Low	Low	Intermediate	Additional file 4: Figure S3
Assay evaluation		Intermediate	Worst	Intermediate	Best

ISSN: 2730-6844