title: LNA-based in situ hybridization detection of mRNAs in embryos.
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.
authors.affiliation: Molecular Cardiovascular Research Program, Department of Cellular and Molecular Medicine, University of Arizona, Tucson, AZ, 85724, USA.
authors.forename: Diana K
authors.initials: DK
authors.lastname: Darnell
authors.forename: Parker B
authors.initials: PB
authors.lastname: Antin
mesh_terms: D000818:Animals; D002642:Chick Embryo / Q000378:metabolism*; D015342:DNA Probes / Q000032:analysis* / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D017403:In Situ Hybridization / Q000379:methods*; D009841:Oligonucleotides / Q000032:analysis* / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics
chemical_list: D015342:DNA Probes; D009841:Oligonucleotides; D012333:RNA, Messenger; C477371:locked nucleic acid