Sensitive and specific detection of microRNAs by northern blot analysis using LNA-modified oligonucleotide probes.

Anna Válóczi, Csaba Hornyik, Nóra Varga, J. Burgyán, Sakari Kauppinen, Z. Havelda

Research output: Contribution to journalArticle

545 Citations (Scopus)

Abstract

We describe here a new method for highly efficient detection of microRNAs by northern blot analysis using LNA (locked nucleic acid)-modified oligonucleotides. In order to exploit the improved hybridization properties of LNA with their target RNA molecules, we designed several LNA-modified oligonucleotide probes for detection of different microRNAs in animals and plants. By modifying DNA oligonucleotides with LNAs using a design, in which every third nucleotide position was substituted by LNA, we could use the probes in northern blot analysis employing standard end-labelling techniques and hybridization conditions. The sensitivity in detecting mature microRNAs by northern blots was increased by at least 10-fold compared to DNA probes, while simultaneously being highly specific, as demonstrated by the use of different single and double mismatched LNA probes. Besides being highly efficient as northern probes, the same LNA-modified oligonucleotide probes would also be useful for miRNA in situ hybridization and miRNA expression profiling by LNA oligonucleotide microarrays.

Original languageEnglish
JournalNucleic Acids Research
Volume32
Issue number22
Publication statusPublished - Dec 2004

Fingerprint

Oligonucleotide Probes
MicroRNAs
Northern Blotting
Oligonucleotides
Nucleic Acid Probes
DNA Probes
locked nucleic acid
Oligonucleotide Array Sequence Analysis
In Situ Hybridization
Nucleotides
RNA
DNA

ASJC Scopus subject areas

  • Genetics

Cite this

Sensitive and specific detection of microRNAs by northern blot analysis using LNA-modified oligonucleotide probes. / Válóczi, Anna; Hornyik, Csaba; Varga, Nóra; Burgyán, J.; Kauppinen, Sakari; Havelda, Z.

In: Nucleic Acids Research, Vol. 32, No. 22, 12.2004.

Research output: Contribution to journalArticle

@article{ceed2ad914c74ddb92c0a78d78385fb3,
title = "Sensitive and specific detection of microRNAs by northern blot analysis using LNA-modified oligonucleotide probes.",
abstract = "We describe here a new method for highly efficient detection of microRNAs by northern blot analysis using LNA (locked nucleic acid)-modified oligonucleotides. In order to exploit the improved hybridization properties of LNA with their target RNA molecules, we designed several LNA-modified oligonucleotide probes for detection of different microRNAs in animals and plants. By modifying DNA oligonucleotides with LNAs using a design, in which every third nucleotide position was substituted by LNA, we could use the probes in northern blot analysis employing standard end-labelling techniques and hybridization conditions. The sensitivity in detecting mature microRNAs by northern blots was increased by at least 10-fold compared to DNA probes, while simultaneously being highly specific, as demonstrated by the use of different single and double mismatched LNA probes. Besides being highly efficient as northern probes, the same LNA-modified oligonucleotide probes would also be useful for miRNA in situ hybridization and miRNA expression profiling by LNA oligonucleotide microarrays.",
author = "Anna V{\'a}l{\'o}czi and Csaba Hornyik and N{\'o}ra Varga and J. Burgy{\'a}n and Sakari Kauppinen and Z. Havelda",
year = "2004",
month = "12",
language = "English",
volume = "32",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "22",

}

TY - JOUR

T1 - Sensitive and specific detection of microRNAs by northern blot analysis using LNA-modified oligonucleotide probes.

AU - Válóczi, Anna

AU - Hornyik, Csaba

AU - Varga, Nóra

AU - Burgyán, J.

AU - Kauppinen, Sakari

AU - Havelda, Z.

PY - 2004/12

Y1 - 2004/12

N2 - We describe here a new method for highly efficient detection of microRNAs by northern blot analysis using LNA (locked nucleic acid)-modified oligonucleotides. In order to exploit the improved hybridization properties of LNA with their target RNA molecules, we designed several LNA-modified oligonucleotide probes for detection of different microRNAs in animals and plants. By modifying DNA oligonucleotides with LNAs using a design, in which every third nucleotide position was substituted by LNA, we could use the probes in northern blot analysis employing standard end-labelling techniques and hybridization conditions. The sensitivity in detecting mature microRNAs by northern blots was increased by at least 10-fold compared to DNA probes, while simultaneously being highly specific, as demonstrated by the use of different single and double mismatched LNA probes. Besides being highly efficient as northern probes, the same LNA-modified oligonucleotide probes would also be useful for miRNA in situ hybridization and miRNA expression profiling by LNA oligonucleotide microarrays.

AB - We describe here a new method for highly efficient detection of microRNAs by northern blot analysis using LNA (locked nucleic acid)-modified oligonucleotides. In order to exploit the improved hybridization properties of LNA with their target RNA molecules, we designed several LNA-modified oligonucleotide probes for detection of different microRNAs in animals and plants. By modifying DNA oligonucleotides with LNAs using a design, in which every third nucleotide position was substituted by LNA, we could use the probes in northern blot analysis employing standard end-labelling techniques and hybridization conditions. The sensitivity in detecting mature microRNAs by northern blots was increased by at least 10-fold compared to DNA probes, while simultaneously being highly specific, as demonstrated by the use of different single and double mismatched LNA probes. Besides being highly efficient as northern probes, the same LNA-modified oligonucleotide probes would also be useful for miRNA in situ hybridization and miRNA expression profiling by LNA oligonucleotide microarrays.

UR - http://www.scopus.com/inward/record.url?scp=16544365954&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=16544365954&partnerID=8YFLogxK

M3 - Article

VL - 32

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 22

ER -