Abstract

Deoxynucleoside 5′-triphosphates (dNTPs) are the molecular building blocks for DNA synthesis, and their balanced concentration in the cell is fundamental for health. dNTP imbalance can lead to genomic instability and other metabolic disturbances, resulting in devastating mitochondrial diseases. The accurate and efficient measurement of dNTPs from different biological samples and cellular compartments is vital to understand the mechanisms behind these diseases and develop and scrutinize their possible treatments. This chapter describes an update on the most recent development of the traditional radiolabeled polymerase extension method and its adaptation for the measurement of whole-cell and mitochondrial dNTP pools from cultured cells and tissue samples. The solid-phase reaction setting enables an increase in efficiency, accuracy, and measurement scale.

Keywords

dNTP; Nucleotide pools; mtDNA; Mitochondrial DNA depletion syndrome; Solid-phase detection

Published in

Methods in Molecular Biology
2021, number: Volume 2276, pages: 143–151
Book title: Mitochondrial medicine
ISBN: 978-1-0716-1265-1, eISBN: 978-1-0716-1266-8
Publisher: HUMANA PRESS INC

Authors' information