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Extracting and measuring dNTP pools in saccharomyces cerevisiae

  • SUNY Buffalo

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

5 Scopus citations

Abstract

Regulation of dNTP pools in an intracellular environment is not only vital for DNA replication but also plays a major role in maintaining genomic stability. Ribonucleotide reductase (RNR) catalyzes the rate-limiting step in dNTP synthesis and altered regulation of RNR leads to imbalanced dNTP pools. Increased dNTP levels are mutagenic and have the potential to interfere with pathways that are involved in DNA replication, repair and DNA damage control. However, the mechanisms through which altered dNTP pools affect these pathways are poorly understood. Nonetheless, altered dNTP pools have been identified in a number of cellular contexts, including cancer. In order to interpret and analyze the effects of altered dNTP pools, we need quantitative information about dNTP pools in different genetic and environmental contexts in vivo. Here we describe a high-throughput fluorescence-based assay that uses a qPCR-based approach to quantify dNTP levels for use with Saccharomyces cerevisiae extracts.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages103-127
Number of pages25
DOIs
StatePublished - 2019

Publication series

NameMethods in Molecular Biology
Volume1999
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Deoxnucleotide triphosphate
  • DNA replication
  • dNTP
  • Genome instability
  • Mutagenesis
  • Quantitative PCR
  • Saccharomyces cerevisiae

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