Gene expression analysis in cartilage by in situ hybridization.
In situ hybridization allows detection and localization of specific nucleic acid sequences directly within a cell or tissue. We present an in situ hybridization protocol using double-stranded DNA or single-stranded RNA probes labeled with [32P] to localize and visualize the temporal and spatial distribution of cartilage-characteristic mRNAs. Probes labeled with this high-energy isotope provide good resolution at the tissue level with relatively low background; as a result of the probes that can be obtained that have a higher specificity to emulsion activity, very short exposure times are required.
Mallein-Gerin
Frédéric
F
Laboratory of Biology and Engineering of Cartilege, CNRS UMR 5086, Institut de Biologie et Chimie des Protéines, Lyon, France.
Gouttenoire
Jérôme
J
D000818:Animals; D002356:Cartilage / Q000737:chemistry / Q000378:metabolism*; D024043:Collagen Type II / Q000235:genetics; D024121:Collagen Type X / Q000235:genetics; D015342:DNA Probes / Q000737:chemistry; D015870:Gene Expression / Q000235:genetics; D017403:In Situ Hybridization / Q000379:methods*; D007867:Leg Bones / Q000737:chemistry; D051379:Mice; D015347:RNA Probes / Q000737:chemistry; D012333:RNA, Messenger / Q000032:analysis*
D024043:Collagen Type II; D024121:Collagen Type X; D015342:DNA Probes; D015347:RNA Probes; D012333:RNA, Messenger