Bio-Rad Laboratories has produced a technical paper (bulletin 5452A) that describes the effect of RNA degradation on data quality in quantitative PCR and microarray experiments
The paper describes using automated electrophoresis to determine RNA quality prior to gene expression analysis.
Quantitative expression levels, when determined by qPCR, decrease as RNA becomes increasingly degraded.
This can lead to erroneous conclusions regarding the levels of gene expression when comparing samples that are degraded to different extents.
Genes with high levels of expression can still be characterised but degradation compromises the ability to detect differences in genes that are expressed at low levels.
Bio-Rad's Experion automated electrophoresis system is an effective method to determine the quality and quantity of RNA in gene expression analysis experiments.
The Experion system accurately quantitates and evaluates the integrity of samples using as little as 25ng total RNA, which is several hundred times less material than that required for gel electrophoresis.
Once RNA quantity and quality have been established, a researcher can determine whether or not to move forward with additional experiments, knowing to what degree the results may be compromised.
The six-page paper provides a detailed introduction, methods description, results, and a discussion, supported by numerous graphs, tables, figures, and electropherograms.
Bulletin 5452A is available either from a local Bio-Rad sales office or it may be downloaded.
