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Optimization of DamID for use in primary cultures of mouse hepatocytes.
Gatticchi, Leonardo; de Las Heras, Jose I; Roberti, Rita; Schirmer, Eric C.
Afiliación
  • Gatticchi L; Department of Experimental Medicine, University of Perugia, Perugia, Italy.
  • de Las Heras JI; Wellcome Centre for Cell Biology, University of Edinburgh, Edinburgh, UK.
  • Roberti R; Department of Experimental Medicine, University of Perugia, Perugia, Italy.
  • Schirmer EC; Wellcome Centre for Cell Biology, University of Edinburgh, Edinburgh, UK. Electronic address: e.schirmer@ed.ac.uk.
Methods ; 157: 88-99, 2019 03 15.
Article en En | MEDLINE | ID: mdl-30445179
DamID, a method to identify DNA associating with a particular protein, was originally developed for use in immortalized tissue culture lines. The power of this technique has led to its adaptation for a number of additional systems. Here we report adaptations for its use in primary cells isolated from rodents with emphasis on the challenges this presents. Specifically, we present several modifications that allow the method to be performed in mouse acutely isolated primary hepatocytes while seemingly maintaining tissue genome architecture. We also describe the downstream bioinformatic analysis necessary to identify LADs and discuss some of the parameters and their effects with regards to the sensitivity of the method.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: ADN / Cromatina / Lamina Tipo B / Cultivo Primario de Células Límite: Animals Idioma: En Revista: Methods Asunto de la revista: BIOQUIMICA Año: 2019 Tipo del documento: Article País de afiliación: Italia Pais de publicación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: ADN / Cromatina / Lamina Tipo B / Cultivo Primario de Células Límite: Animals Idioma: En Revista: Methods Asunto de la revista: BIOQUIMICA Año: 2019 Tipo del documento: Article País de afiliación: Italia Pais de publicación: Estados Unidos