1Department of Physiology, University of Oklahoma College of Medicine, 2Department of Geriatric Medicine, University of Oklahoma College of Medicine
Bisulfite amplicon sequencing (BSAS) is a method for quantifying cytosine methylation in targeted genomic regions of interest. This method uses bisulfite conversion paired with PCR amplification of target regions prior to next-generation sequencing to produce absolute quantitation of DNA methylation at a base-specific level.
Published February 24, 2015. Keywords: Molecular Biology, Epigenetics, DNA methylation, next-generation sequencing, bioinformatics, gene expression, cytosine, CpG, gene expression regulation
1Diagenode S.A., 2Diagenode Inc.
Methods for mapping in vivo protein-DNA interactions are becoming crucial for every aspect of genomic research but they are laborious, costly, and time consuming. Here a commercially available robotic liquid handling system that automates chromatin immunoprecipitation for mapping in vivo protein-DNA interactions with limited amounts of cells is presented.
Published December 10, 2014. Keywords: Molecular Biology, Automation, chromatin immunoprecipitation, low DNA amounts, histone antibodies, sequencing, library preparation
1Department of Basic Sciences, Mississippi State University College of Veterinary Medicine
The isolation of individual dopamine neurons or the ventral tegmental area with direct or indirect immunohistochemistry is demonstrated using laser capture microdissection. Parameters for isolation of tissue from a glass slide using an infrared laser and from membrane slides using the combination of an infrared and ultraviolet laser are discussed.
Published February 6, 2015. Keywords: Neuroscience, Laser capture microdissection, dopamine neuron, Immunohistochemistry, Tyrosine hydroxylase, Ventral tegmental area, PEN membrane glass slide.
1Virus and Centromere Team, Centre de Génétique et Physiologie Moléculaire et Cellulaire, CNRS UMR 5534, 2Université de Lyon 1, 3Laboratoire d'excellence, LabEX DEVweCAN, 4Institut de Virologie Moléculaire et Structurale, CNRS UPR 3296, 5Centre de Recherche en Cancérologie de Lyon, INSERM U1052, CNRS UMR 5286
We established a fluorescent in situ hybridization protocol for the detection of a persistent DNA virus genome within tissue sections of animal models. This protocol enables studying infection process by codetection of the viral genome, its RNA products, and viral or cellular proteins within single cells.
Published January 23, 2014. Keywords: Neuroscience, Life Sciences (General), Virology, Herpes Simplex Virus (HSV), Latency, In situ hybridization, Nuclear organization, Gene expression, Microscopy
1Department of Integrative Oncology, BC Cancer Research Centre, 2Interdisciplinary Oncology Program, University of British Columbia - UBC, 3Photography/Video Production, Multi-Media Services, BC Cancer Agency, 4Department of Pathology and Laboratory Medicine, University of British Columbia - UBC
This video demonstrates the protocol for DNA extraction from formalin-fixed paraffin-embedded material. This is a multi-day procedure in which tissue sections are deparaffinized with xylene, rehydrated with ethanol and treated with proteinase K to purify and isolate DNA for subsequent gene-specific or genome-wide analysis.
Published March 26, 2011. Keywords: Genetics, DNA extraction, paraffin embedded tissue, phenol:chloroform extraction, genetic analysis, epigenetic analysis
1Strategic Research Center for Stem Cell Biology and Cell Therapy, University of Lund, 2Department of Cardiology Lund University Hospital, University of Lund
Cardiac nuclei are isolated via density sedimentation and immunolabeled with antibodies against pericentriolar material 1 (PCM-1) to identify and sort cardiomyocyte nuclei by flow cytometry.
Published July 10, 2012. Keywords: Medicine, Stem Cell Biology, Cardiology, Physiology, Tissue Engineering, cardiomyocyte, post mortem, nuclei isolation, flow cytometry, pericentriolar material 1, PCM-1