Abstract
Here, we described a method for carrying out nonradioactive in situ hybridization to detect mRNA transcripts in cryostat sections of mouse bone using the CryoJane® Tape-Transfer System and digoxigenin (DIG)-labeled riboprobes.
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Acknowledgments
We thank Dr. M. John (Novartis Institutes for BioMedical Research, Basel, Switzerland) and Prof. E. Schipani (Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA) for providing the DNA constructs for generation of type 1 collagen alpha 1 and type 10 collagen riboprobes. We are grateful to Profs. D. W. Rowe and I. Kalajzic (University of Connecticut Health Center, Farmington, CT, USA) as well as Prof. S. E. Harris (University of Texas Health Science Center at San Antonio, San Antonio, TX, USA) for advice on fixation and decalcification times, as well as cryo-sectioning with the CryoJane® Tape-Transfer System and ISH on skeletal tissue.
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Kramer, I., Salie, R., Susa, M., Kneissel, M. (2012). Studying Gene Expression in Bone by In Situ Hybridization. In: Helfrich, M., Ralston, S. (eds) Bone Research Protocols. Methods in Molecular Biology, vol 816. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-415-5_20
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DOI: https://doi.org/10.1007/978-1-61779-415-5_20
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