Analysis of Liver Connexin Expression Using Reverse Transcription Quantitative Real-Time Polymerase Chain Reaction

Methods Mol Biol. 2016:1437:1-19. doi: 10.1007/978-1-4939-3664-9_1.

Abstract

Although connexin production is mainly regulated at the protein level, altered connexin gene expression has been identified as the underlying mechanism of several pathologies. When studying the latter, appropriate methods to quantify connexin RNA levels are required. The present chapter describes a well-established reverse transcription quantitative real-time polymerase chain reaction procedure optimized for analysis of hepatic connexins. The method includes RNA extraction and subsequent quantification, generation of complementary DNA, quantitative real-time polymerase chain reaction, and data analysis.

Keywords: Connexins; Minimum Information for publication of Quantitative real-time PCR Experiments; RNA extraction; Reverse transcription.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Connexins / metabolism*
  • Guidelines as Topic
  • Humans
  • Liver / metabolism*
  • Mice
  • RNA / metabolism
  • Real-Time Polymerase Chain Reaction / methods*
  • Reverse Transcriptase Polymerase Chain Reaction / methods*

Substances

  • Connexins
  • RNA