Factor correction as a tool to eliminate between-session variation in replicate experiments: application to molecular biology and retrovirology

• Main Authors: Das Atze T, Schoneveld Onard JLM, Thygesen Helene H, Ruijter Jan M, Berkhout Ben, Lamers Wouter H
• Format: Article
• Language: English
• Published: BMC 2006-01-01
• Series: Retrovirology
• Online Access: http://www.retrovirology.com/content/3/1/2

<p>Abstract</p> <p>Background</p> <p>In experimental biology, including retrovirology and molecular biology, replicate measurement sessions very often show similar proportional differences between experimental conditions, but different absolute values, even though the measurements were presumably carried out under identical circumstances. Although statistical programs enable the analysis of condition effects despite this replication error, this approach is hardly ever used for this purpose. On the contrary, most researchers deal with such between-session variation by normalisation or standardisation of the data. In normalisation all values in a session are divided by the observed value of the 'control' condition, whereas in standardisation, the sessions' means and standard deviations are used to correct the data. Normalisation, however, adds variation because the control value is not without error, while standardisation is biased if the data set is incomplete.</p> <p>Results</p> <p>In most cases, between-session variation is multiplicative and can, therefore, be removed by division of the data in each session with a session-specific correction factor. Assuming one level of multiplicative between-session error, unbiased session factors can be calculated from all available data through the generation of a between-session ratio matrix. Alternatively, these factors can be estimated with a maximum likelihood approach. The effectiveness of this correction method, dubbed "factor correction", is demonstrated with examples from the field of molecular biology and retrovirology. Especially when not all conditions are included in every measurement session, factor correction results in smaller residual error than normalisation and standardisation and therefore allows the detection of smaller treatment differences. Factor correction was implemented into an easy-to-use computer program that is available on request at: <email>[e-mail]?subject=factor</email>.</p> <p>Conclusion</p> <p>Factor correction is an effective and efficient way to deal with between-session variation in multi-session experiments.</p>