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Garvan Institute of Medical Research

3 ARTICLES PUBLISHED IN JoVE

Biology

DNA Methylation: Bisulphite Modification and Analysis

Kate Patterson^*1, Laura Molloy^*1, Wenjia Qu¹, Susan Clark^1,2

¹Epigenetics Group, Cancer Research Program, Garvan Institute of Medical Research, ²St Vincent's Clinical School, University of NSW

The gold standard for DNA methylation analysis is genomic sequencing of bisulphite converted DNA. This method takes advantage of the increased sensitivity of cytosine compared with 5-methylcytosine (5-MeC) to bisulphite deamination under acidic conditions. Unmethylated cytosines can be distinguished from methylated cytosines after PCR amplification of the target genomic DNA.

Neuroscience

Preparation of an Awake Mouse for Recording Neural Responses and Injecting Tracers

Michael A. Muniak^1,2, Zachary M. Mayko³, David K. Ryugo^2,4, Christine V. Portfors³

¹Department of Neuroscience, Johns Hopkins University, ²Garvan Institute of Medical Research, ³School of Biological Sciences, Washington State University, ⁴Department of Otolaryngology-HNS, Johns Hopkins University

Electrophysiological characterization of neuronal responses is important for understanding brain function and for guiding the placement of dyes for pathway tracing. However, many studies are performed in anesthetized animals. To understand brain function without anesthetics, we developed a method to record neuronal response properties and inject dyes in awake mouse.

Immunology and Infection

Dissecting Multi-protein Signaling Complexes by Bimolecular Complementation Affinity Purification (BiCAP)

Jordan F. Hastings¹, Jeremy Z.R. Han¹, Robert F. Shearer^1,2, Sean P. Kennedy^1,3, Mary Iconomou^1,4, Darren N. Saunders⁵, David R. Croucher^1,6,7

¹The Kinghorn Cancer Centre, Garvan Institute of Medical Research, ²Ubiquitin Signaling Group, Protein Signaling Program, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, ³RCSI Molecular Medicine, Royal College of Surgeons in Ireland, ⁴Department of Epigenetics, Max Planck Institute of Immunobiology and Epigenetics, ⁵School of Medical Sciences, University of New South Wales, ⁶St Vincent's Hospital Clinical School, University of New South Wales, ⁷School of Medicine and Medical Science, University College Dublin

This manuscript describes the protocol for Bimolecular Complementation Affinity Purification (BiCAP). This novel method facilitates the specific isolation and downstream proteomic characterization of any two interacting proteins, while excluding un-complexed individual proteins as well as complexes formed with competing binding partners.