Novel method to ascertain chromatin accessibility at specific genomic loci from frozen brain homogenates and laser capture microdissected defined cells.
Neuroepigenetics
; 6: 1-9, 2016 Jun.
Article
em En
| MEDLINE
| ID: mdl-27158594
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 an in vitro preparation. Although we demonstrate the use of this method in brain tissue we also recognize its applicability to other tissue types.
Texto completo:
1
Base de dados:
MEDLINE
Idioma:
En
Revista:
Neuroepigenetics
Ano de publicação:
2016
Tipo de documento:
Article
País de afiliação:
Estados Unidos