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.
Laboratory of Biology and Engineering of Cartilege, CNRS UMR 5086, Institut de Biologie et Chimie des Protéines, Lyon, France.
Frédéric
F
Mallein-Gerin
Jérôme
J
Gouttenoire
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