Biotechnology Letters

, 32:35

Validation of extraction methods for total RNA and miRNA from bovine blood prior to quantitative gene expression analyses

  • Andrea Hammerle-Fickinger
  • Irmgard Riedmaier
  • Christiane Becker
  • Heinrich H. D. Meyer
  • Michael W. Pfaffl
  • Susanne E. Ulbrich
Original Research Paper

DOI: 10.1007/s10529-009-0130-2

Cite this article as:
Hammerle-Fickinger, A., Riedmaier, I., Becker, C. et al. Biotechnol Lett (2010) 32: 35. doi:10.1007/s10529-009-0130-2

Abstract

The benefit and precision of blood diagnosis by quantitative real-time PCR (qPCR) is limited by sampling procedures and RNA extraction methods. We have compared five different RNA extraction protocols from bovine blood regarding RNA and miRNA yield, quality, and most reproducible data in the qRT-PCR with the lowest point of quantification. Convincing results in terms of highest quantity, quality, and best performance for mRNA qPCR were obtained by leukocyte extraction following blood lysis as well as extraction of PAXgene stabilized blood. The best microRNA qPCR results were obtained for samples extracted by the leukocyte extraction method.

Keywords

Blood Bovine miRNA Molecular diagnosis mRNA qPCR 

Supplementary material

10529_2009_130_MOESM1_ESM.pdf (39 kb)
Supplementary material 1 (DOC 40 kb)

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Andrea Hammerle-Fickinger
    • 1
  • Irmgard Riedmaier
    • 1
  • Christiane Becker
    • 1
  • Heinrich H. D. Meyer
    • 1
  • Michael W. Pfaffl
    • 1
  • Susanne E. Ulbrich
    • 1
  1. 1.Physiology WeihenstephanTechnische Universitaet MuenchenFreisingGermany

Personalised recommendations