Advancing Epigenetics Towards Systems Biology

Quantitative Chromosome Conformation Capture (3C-qPCR) (Prot 65)

Vuthy Ea1, Thierry Forné1, Franck Court1,2

Introduction

Here we describe a modified 3C-qPCR protocol with four-fold increased efficiency of the 3C assay compared to the initially published protocols (Ref.1-4). Classical SybrGreen technique can then be used, instead of TaqMan probes, to quantify by quantitative PCR relevant 3C chimerical ligation products. It is optimized for nuclei preparation as starting material, coming from fresh tissues or cultured cells. In our hands, it was successfully used on human and mouse samples.

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Vuthy Ea1, Thierry Forné1, Franck Court1,2

1 Institut de Génétique Moléculaire, UMR5535 CNRS University of Montpellier, France
2 Present address: Inserm UMR1103, CNRS UMR6293, F-63001 Clermont-Ferrand, France and Clermont Université, Université d'Auvergne, Laboratoire GReD, BP 10448, F-63000 Clermont-Ferrand, France.

Corresponding author: Franck Court
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Franck Court