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

Methods Mol Biol. 2014:1160:43-53. doi: 10.1007/978-1-4939-0733-5_5.

Abstract

RNA quality control is a crucial step in guaranteeing integer nondegraded RNA and receiving meaningful results in gene expression profiling 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.

MeSH terms

  • Animals
  • Cattle
  • Electrophoresis, Capillary
  • Gene Expression Profiling / methods*
  • Mice
  • MicroRNAs / genetics*
  • Microfluidic Analytical Techniques
  • Polymerase Chain Reaction
  • RNA, Messenger / genetics*

Substances

  • MicroRNAs
  • RNA, Messenger