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Detection of Small Noncoding RNAs by In Situ Hybridization Using Probes of 2′-O-Methyl RNA + LNA

Publikation: Bidrag til tidsskriftTidsskriftartikel

  • Martin Jensen Søe
  • Martin Dufva
  • Kim Holmstrøm
In situ hybridization is a powerful method to provide information about contextual distribution and cellular origin of nucleic acids, e.g., in formalin-fixed paraffin-embedded (FFPE) samples of tissue. Particularly the recently discovered classes of noncoding RNA (ncRNA) including endo-siRNAs and microRNAs require such a technique to enable their study and visualization in natural contexts, and in the last decade, many advances have been made, increasing our ability to specifically detect small ncRNAs. One of the key developments has been the demonstration of the superiority of using locked nucleic acid (LNA)-modified DNA probes for the detection of ncRNA in tissue. Here, we describe an alternative in situ hybridization protocol employing oligonucleotide probes consisting of combinations of LNA and 2´-O-methyl RNAs that under optimized hybridization buffer conditions can provide a highly sensitive assay performance with only 1 h hybridization time.
OriginalsprogEngelsk
TidsskriftMethods in Molecular Biology
Vol/bind1173
Sider (fra-til)113-121
ISSN1064-3745
DOI
StatusUdgivet - 18 jun. 2014
Eksternt udgivetJa

ID: 117196737