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.
MeSH terms
-
Animals
-
Bone and Bones / cytology*
-
Cell Line, Tumor
-
DNA / genetics
-
DNA / isolation & purification
-
Digoxigenin / chemistry
-
Gene Expression Profiling / instrumentation
-
Gene Expression Profiling / methods
-
Histocytological Preparation Techniques / instrumentation
-
Histocytological Preparation Techniques / methods
-
Humans
-
In Situ Hybridization / instrumentation
-
In Situ Hybridization / methods*
-
Mice
-
Mice, Nude
-
MicroRNAs / genetics
-
MicroRNAs / isolation & purification
-
Oligonucleotides / chemistry
-
RNA, Messenger / genetics
-
RNA, Messenger / isolation & purification
-
Real-Time Polymerase Chain Reaction / instrumentation
-
Real-Time Polymerase Chain Reaction / methods*
-
Reverse Transcriptase Polymerase Chain Reaction / instrumentation
-
Reverse Transcriptase Polymerase Chain Reaction / methods*
-
Xenograft Model Antitumor Assays
Substances
-
MicroRNAs
-
Oligonucleotides
-
RNA, Messenger
-
locked nucleic acid
-
DNA
-
Digoxigenin