Analysis of replicative polymerase usage by ribonucleotide incorporation

Andrea Keszthelyi, Izumi Miyabe, Katie Ptasińska, Yasukazu Daigaku, Karel Naiman, Antony M. Carr

研究成果: Chapter

抄録

Mapping the usage of replicative DNA polymerases has previously proved to be technically challenging. By exploiting mutant polymerases that incorporate ribonucleotides into the DNA with a significantly higher proficiency than their wild-type counterparts, we and others have developed methods that can identify what proportion of each DNA strand (i.e., the Watson and Crick strands) is replicated by a specific DNA polymerase. The incorporation of excess ribonucleotides by a mutated polymerase effectively marks, in each individual cells, the DNA strand that is replicated by that specific mutated polymerase. Changes to DNA polymerase usage can be examined at specific loci by Southern blot analysis while a global analysis of polymerase usage can be achieved by applying next-generation sequencing. This genome-wide data also provides a direct measure of replication origin efficiency and can be used to indirectly calculate replication timing.

本文言語English
ホスト出版物のタイトルMethods in Molecular Biology
出版社Humana Press Inc.
ページ239-259
ページ数21
DOI
出版ステータスPublished - 2018 1 1

出版物シリーズ

名前Methods in Molecular Biology
1672
ISSN(印刷版)1064-3745

ASJC Scopus subject areas

  • 分子生物学
  • 遺伝学

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