Isolation of high spectral quality RNA using run-on gene transcription; application to gene expression profiling of human brain

Jian Guo Cui, Yuhai Zhao, Walter J. Lukiw

Research output: Contribution to journalArticle

18 Scopus citations

Abstract

1. Gene expression studies of human brain tissues in both health and disease require the use of intact, high spectral quality messenger RNA (mRNA). Short post-mortem tissues, when adequately controlled, provide a valuable primary source of brain mRNA. 2. Typical mRNA isolation methodologies rely on total RNA extraction from whole cells or tissues. Here we report that as an alternative approach, nuclei can be isolated from the same tissues and programmed for run-on gene transcription to generate enriched mRNA fractions. 3. This novel technique represents a significant improvement over previous methods and provides high quality, high yield mRNA samples suitable for downstream applications that include cloning, sequencing and gene expression studies using DNA array technologies.

Original languageEnglish (US)
Pages (from-to)789-794
Number of pages6
JournalCellular and molecular neurobiology
Volume25
Issue number3-4
DOIs
StatePublished - Jun 1 2005

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Keywords

  • Alzheimer's disease (AD)
  • Brain transcription
  • Messenger RNA
  • Micro-machines
  • Nanotechnology
  • RNA polymerase II
  • Run-on gene transcription

ASJC Scopus subject areas

  • Neuroscience(all)
  • Clinical Biochemistry
  • Cell Biology
  • Genetics

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