Dramatically improved RNA in situ hybridization signals using LNA-modified probes

  1. RUNE THOMSEN1,
  2. PETER STEIN NIELSEN2, and
  3. TORBEN HEICK JENSEN1
  1. 1Centre for mRNP Biogenesis and Metabolism, Department of Molecular Biology, Aarhus University, DK-8000 Aarhus C, Denmark
  2. 2Department of Functional Genomics, Exiqon, DK-2950 Vedbaek, Denmark

Abstract

In situ detection of RNA by hybridization with complementary probes is a powerful technique. Probe design is a critical parameter in successful target detection. We have evaluated the efficiency of fluorescent DNA oligonucleotides modified to contain locked nucleic acid (LNA) residues. This increases the thermal stability of hybrids formed with RNA. The LNA-based probes detect specific RNAs in fixed yeast cells with an efficiency far better than conventional DNA oligonucleotide probes of the same sequence. Using this probe design, we were also able to detect poly(A)+ RNA accumulation within the nucleus/ nucleolus of wild-type cells. LNA-based probes should be readily applicable to a diverse array of cells and tissue samples.

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  1. Published in Advance September 21, 2005, doi: 10.1261/rna.2139705 RNA 2005. 11: 1745-1748 Copyright 2005 by RNA Society

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