mRNA and microRNA purity and integrity: The key to success in expression profiling

Benedikt Kirchner, Vijay Paul, Irmgard Riedmaier, Michael W. Pfaffl

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

RNA quality control is a crucial step in guaranteeing integer nondegraded RNA and receiving meaningful results in gene expression profi ling experiments, using micro-array, RT-qPCR (Reverse-Transcription quantitative PCR), or Next-Generation-Sequencing by RNA-Seq or small-RNA Seq. Therefore, assessment of RNA integrity and purity is very essential prior to gene expression analysis of sample RNA to ensure the accuracy of any downstream applications. RNA samples should be nondegraded or fragmented and free of protein, genomic DNA, nucleases, and enzymatic inhibitors. Herein we describe the current state-of-the-art RNA quality assessment by combining UV/Vis spectrophotometry and microfluidic capillary electrophoresis.

Original languageEnglish
Pages (from-to)43-53
Number of pages11
JournalMethods in molecular biology (Clifton, N.J.)
Volume1160
DOIs
StatePublished - 2014

Keywords

  • Denaturing gel electrophoresis
  • MIQE
  • MicroRNA (miRNA)
  • Microfluidic capillary electrophoresis
  • RNA integrity
  • RNA purity
  • UV/Vis spectrophotometer
  • mRNA
  • qPCR

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