LNA-based in situ hybridization detection of mrnas in embryos

Diana K. Darnell, Parker B Antin

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

In situ hybridization (ISH) in embryos allows the visualization of specific RNAs as a readout of gene expression during normal development or after experimental manipulations. ISH using short DNA probes containing locked nucleic acid nucleotides (LNAs) holds the additional advantage of allowing the detection of specific RNA splice variants or of closely related family members that differ in only short regions, creating new diagnostic and detection opportunities. Here we describe methods for using short (14–24 nt) DNA probes containing LNA nucleotides to detect moderately to highly expressed RNAs in whole chick embryos during the first 5 days of embryonic development. The protocol is easily adaptable for use with embryos of other vertebrate species.

Original languageEnglish (US)
Pages (from-to)69-76
Number of pages8
JournalMethods in molecular biology (Clifton, N.J.)
Volume1211
DOIs
StatePublished - 2014
Externally publishedYes

Fingerprint

In Situ Hybridization
Embryonic Structures
DNA Probes
RNA
Nucleotides
Chick Embryo
Embryonic Development
Vertebrates
Gene Expression
locked nucleic acid

Keywords

  • Chicken embryo
  • Gallus gallus
  • In situ hybridization
  • LNA
  • Locked nucleic acids

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Medicine(all)

Cite this

LNA-based in situ hybridization detection of mrnas in embryos. / Darnell, Diana K.; Antin, Parker B.

In: Methods in molecular biology (Clifton, N.J.), Vol. 1211, 2014, p. 69-76.

Research output: Contribution to journalArticle

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