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Microsoft Excel was used to calculate standard deviations (SD) and statistically significant differences between samples using the two-tailed Student's t-test.
Additionally, expression of the 18S ribosomal subunit was also assessed to allow for normalization of total RNA levels between samples using the 2−ΔΔCT method [38].
A principal component analysis was conducted to detect overall differences between samples using the STAMP programme.
UniFrac is a method to calculate a distance measure between samples using the phylogenetic trees.
The data was normalised across library sizes, between samples using the trimmed mean of M-values (TMM) normalization method [ 70].
The data were normalised across library size between samples using the trimmed mean of M-values normalization method [ 58] (Additional file 1c).
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Seriation analyses along the investigated time line were conducted to test for the similarity of ARISA patterns between consecutive samples using the Spearman rank correlation method.
The geometrical parameters of vertebrae were statistically compared between different samples using the T-test for two independent samples (SPSS 14.0).
Keliadegraph was used to plot the distribution of intensities and to determine the significance of any variability between the samples using the Students t-test.
Indeed, the high degree of concordance between the samples using the Spearman Rank Correlation shows that for the most part, these two methods are comparable.
Referring to "The significance of digital gene expression profiles [ 63]", BGI have developed a strict algorithm to identify differentially expressed genes between two samples using the Poisson model.
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