title: A rapid and convenient method to prepare DIG-labelled RNA probes for use in non-radioactive in situ hybridization.
abstract: We describe here the use of PCR-generated templates incorporating T3 polymerase sites in order to prepare digoxigenin (DIG)-labelled cRNA probes against any gene of known sequence. This method was applied to the preparation of probes specific for chicken glyceraldehyde-3-phosphate dehydrogenase messenger RNAs and we demonstrate that such probes can be used for in situ hybridization (ISH). This technique therefore represents a rapid and convenient means to prepare DIG-labelled cRNA probes for use in a non-radioactive ISH. It adds speed and convenience of probe preparation to the previously described advantages of non-radioactive detection techniques.
authors.affiliation: Département de Biologie Moléculaire et Cellulaire, Ecole Normale Supérieure de Lyon, France.
authors.forename: O
authors.initials: O
authors.lastname: Gandrillon
authors.forename: F
authors.initials: F
authors.lastname: Solari
authors.forename: C
authors.initials: C
authors.lastname: Legrand
authors.forename: P
authors.initials: P
authors.lastname: Jurdic
authors.forename: J
authors.initials: J
authors.lastname: Samarut
mesh_terms: D000818:Animals; D001483:Base Sequence; D002645:Chickens; D004076:Digoxigenin; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D017403:In Situ Hybridization / Q000379:methods*; D008264:Macrophages; D015336:Molecular Probe Techniques; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D018075:RNA, Complementary; D012333:RNA, Messenger / Q000032:analysis
chemical_list: D018075:RNA, Complementary; D012333:RNA, Messenger; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D004076:Digoxigenin