An inter-platform repeatability study investigating real-time amplification of plasmid DNA

BMC Biotechnology

Table 2 Variation attributable to day, plasmid copy number, PCR reaction mix and platform following an ANOVA test using threshold cycle number (Ct) as the variable factor.

	Univariate Tests of Significance for Ct value. Sigma-restricted parameterisation. Effective hypothesis decomposition.
Effect	SS	DF	MS	F	P
Day	96.0	2	48.0	221.4	<0.00001
Plasmid Copies	12919.2	6	2153.2	9932.0	<0.00001
PCR Reaction Mix	75.0	1	75.0	346.2	<0.00001
Platform	700.5	2	350.2	1615.5	<0.00001
Day with Plasmid Copies	53.2	12	4.4	20.5	<0.00001
Day with PCR Reaction Mix	1.9	2	0.9	4.3	0.01442
Plasmid Copies with PCR Reaction Mix	8.5	6	1.4	6.5	<0.00001
Day with Platform	130.8	4	32.7	150.8	<0.00001
Plasmid Copies with Platform	51.0	12	4.2	19.6	<0.00001
PCR Reaction Mix with Platform	1.7	2	0.9	4.0	0.01901
Day with Plasmid Copies with PCR Reaction Mix	14.6	12	1.2	5.6	<0.00001
Day with Plasmid copies with Platform	18.3	24	0.8	3.5	<0.00001
Day with PCR Reaction Mix with Platform	49.6	4	12.4	57.1	<0.00001
Plasmid copies with PCR Reaction Mix with Platform	4.5	12	0.4	1.7	0.06337
Day with Plasmid Copies with PCR Reaction Mix with Platform	11.8	24	0.5	2.3	0.00091
Error	54.4	251	0.2

(Where SS = sum of squares; DF = degrees of freedom; MS = Mean Square; F = ratio of variances; p = probability of obtaining a specific result, given by the null hypothesis).

ISSN: 1472-6750