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Methods Mol Biol. 2019;1914:169-196. doi: 10.1007/978-1-4939-8997-3_9.

Analysis of mRNA, miRNA, and DNA in Bone Cells by RT-qPCR and In Situ Hybridization.

Author information

1
INSERM, UMR1238, Bone Sarcoma and Remodeling of Calcified Tissues, Université de Nantes, Nantes Atlantique Universités, Nantes, France.
2
INSERM, UMR1238, Bone Sarcoma and Remodeling of Calcified Tissues, Université de Nantes, Nantes Atlantique Universités, Nantes, France. francois.lamoureux@univ-nantes.fr.

Abstract

The aim of this chapter is to describe a method used to evaluate gene expression and microRNAs (miRNAs) in bone cells or tissue using Reverse transcription and quantitative Polymerase Chain Reaction (RT-qPCR), and a method to assess chromogenic in situ hybridization (CISH) on Formalin Fixed Paraffin Embedded (FFPE ) mouse bone tissue to detect both DNA and mRNA transcripts using the double digoxigenin (DIG) locked nucleic acid (LNA™) probes .

KEYWORDS:

Bone; Bone sarcoma; Chromogenic in situ hybridization; DNA; Gene expression; Histology; LNA probes; Primers; RT-qPCR; mRNA; miRNA

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