title: Novel method to ascertain chromatin accessibility at specific genomic loci from frozen brain homogenates and laser capture microdissected defined cells.
abstract: We describe a novel method for assessing the "open" or "closed" state of chromatin at selected locations within the genome. This method combines the use of Benzonase, which can digest DNA in the presence of actin, with qPCR to define digested regions. We demonstrate the application of this method in brain homogenates and laser captured cells. We also demonstrate application to selected sites within more than one gene and multiple sites within one gene. We demonstrate the validity of the method by treating cells with valproate, known to render chromatin more permissive, and by comparison with classical digestion with DNase I in anin vitropreparation. Although we demonstrate the use of this method in brain tissue we also recognize its applicability to other tissue types.
authors.affiliation: ASU-Banner Neurodegenerative Disease Research Center, Biodesign Institute and School of Life Sciences, Arizona State University, Tempe, AZ, USA; L.J. Roberts Center for Alzheimer's Research, Banner Sun Health Research Institute, 10515 W Santa Fe Dr, Sun City, AZ 85351, USA.
authors.forename: Elaine
authors.initials: E
authors.lastname: Delvaux
authors.affiliation: ASU-Banner Neurodegenerative Disease Research Center, Biodesign Institute and School of Life Sciences, Arizona State University, Tempe, AZ, USA; L.J. Roberts Center for Alzheimer's Research, Banner Sun Health Research Institute, 10515 W Santa Fe Dr, Sun City, AZ 85351, USA; Department of Psychiatry and Neuropsychology, School for Mental Health and Neuroscience (MHeNS), Faculty of Health, Medicine and Life Sciences, European Graduate School of Neuroscience (EURON), Maastricht University Medical Centre, Maastricht, The Netherlands.
authors.forename: Diego
authors.initials: D
authors.lastname: Mastroeni
authors.affiliation: ASU-Banner Neurodegenerative Disease Research Center, Biodesign Institute and School of Life Sciences, Arizona State University, Tempe, AZ, USA; L.J. Roberts Center for Alzheimer's Research, Banner Sun Health Research Institute, 10515 W Santa Fe Dr, Sun City, AZ 85351, USA.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Nolz
authors.affiliation: ASU-Banner Neurodegenerative Disease Research Center, Biodesign Institute and School of Life Sciences, Arizona State University, Tempe, AZ, USA; L.J. Roberts Center for Alzheimer's Research, Banner Sun Health Research Institute, 10515 W Santa Fe Dr, Sun City, AZ 85351, USA.
authors.forename: Paul D
authors.initials: PD
authors.lastname: Coleman