Open Access

Tracking chromatin states using controlled DNase I treatment and real-time PCR

  • Rui Pires Martins1,
  • Adrian E. Platts1, 2 and
  • Stephen A. Krawetz1, 3Email author
Cellular & Molecular Biology LettersAn International Journal200712:24

DOI: 10.2478/s11658-007-0024-z

Received: 21 February 2007

Accepted: 22 May 2007

Published: 24 June 2007

Abstract

A novel approach to DNase I-sensitivity analysis was applied to examining genes of the spermatogenic pathway, reflective of the substantial morphological and genomic changes that occur during this program of differentiation. A new real-time PCR-based strategy that considers the nuances of response to nuclease treatment was used to assess the nuclease susceptibility through differentiation. Data analysis was automated with the K-Lab PCR algorithm, facilitating the rapid analysis of multiple samples while eliminating the subjectivity usually associated with Ct analyses. The utility of this assay and analytical paradigm as applied to nuclease-sensitivity mapping is presented.

Key words

DNase I-sensitivity Differentiation Spermatogenesis Method

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