title: Evaluation of methods for amplification of picogram amounts of total RNA for whole genome expression profiling.
abstract: BACKGROUND
  For more than a decade, microarrays have been a powerful and widely used tool to explore the transcriptome of biological systems. However, the amount of biological material from cell sorting or laser capture microdissection is much too small to perform microarray studies. To address this issue, RNA amplification methods have been developed to generate sufficient targets from picogram amounts of total RNA to perform microarray hybridisation.
  
  
  RESULTS
  In this study, four commercial protocols for amplification of picograms amounts of input RNA for microarray expression profiling were evaluated and compared. The quantitative and qualitative performances of the methods were assessed. Microarrays were hybridised with the amplified targets and the amplification protocols were compared with respect to the quality of expression profiles, reproducibility within a concentration range of input RNA, and sensitivity. The results demonstrate significant differences between these four methods.
  
  
  CONCLUSIONS
  In our hands, the WT-Ovation pico system proposed by Nugen appears to be the most suitable for RNA amplification. This comparative study will be useful to scientists needing to choose an amplification method to carry out microarray experiments involving samples comprising only a few cells and generating picogram amounts of RNA.
authors.affiliation: Institut Curie, Département de Transfert, 26 Rue d'ULM, F-75248, Paris cedex 05, France. mathieu.clement-ziza@biotec.tu-dresden.de
authors.forename: Mathieu
authors.initials: M
authors.lastname: Clément-Ziza
authors.forename: David
authors.initials: D
authors.lastname: Gentien
authors.forename: Stanislas
authors.initials: S
authors.lastname: Lyonnet
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: Thiery
authors.forename: Claude
authors.initials: C
authors.lastname: Besmond
authors.forename: Charles
authors.initials: C
authors.lastname: Decraene
mesh_terms: D002460:Cell Line; D020869:Gene Expression Profiling; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D012313:RNA / Q000302:isolation & purification*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D012313:RNA