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Table 1 Experimental setup of the four qPCR assays for relative telomere length estimation

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

Reaction conditions    Primers   Standards    Telomere Reference
Assay Platform Type Master mix Duration Tel Ref Batches Dilutions LDR Cq NTC Cq NTC
I ABI Singleplex Commercial 21 CW2006 BA 1998 7 5 4.5-72.0 17.6 34.9 33.5 40.0
II RG Multiplex RC 2009 6 RC 2009 RC 2009 4 4 2.2-60.0 15.7 19.4 39.9 27.9
III RG Singleplex RC 2009 5 RC 2009 RC 2009 4 4 2.2-60.0 14.1 15.5 37.4 36.2
IV RG Multiplex Commercial 6 RC 2009 RC 2009 4 5 4.5-72.0 13.9 27.2 34.3 31.1
  1. Duration = number of days it took to generate the data once assays had been optimized; BA = Bérubé and Aguilar [38]; CW 2006 = Callicot and Womack [52]; RC 2009 = Cawthon [24]; Batches = number of separate qPCR runs (plates); Dilutions = number of dilution steps; LDR = linear dynamic range (ng/reaction); Cq = the average Cq of the serially diluted standard with the lowest amount of DNA; NTC = the lowest Cq value observed for a NTC.