Ribonucleotides (rNTPs) are incorporated into genomic DNA at a relatively high frequency during replication. They have beneficial effects but, if not removed from the chromosomes, increase genomic instability. Here, we describe a fast method to easily estimate the amounts of embedded ribonucleotides into the genome. The protocol described is performed in Saccharomyces cerevisiae and allows us to quantify altered levels of rNMPs due to different mutations in the replicative polymerase ε. However, this protocol can be easily applied to cells derived from any organism.

Measuring the levels of ribonucleotides embedded in genomic DNA / A. Meroni, M. Nava Giulia, S. Sertic, P. Plevani, M. Muzi-Falconi, F. Lazzaro (METHODS IN MOLECULAR BIOLOGY). - In: Genome Instability / [a cura di] M. Muzi-Falconi, G.W. Brown. - [s.l] : Humana Press Inc., 2018. - ISBN 9781493973057. - pp. 319-327 [10.1007/978-1-4939-7306-4_22]

Measuring the levels of ribonucleotides embedded in genomic DNA

A. Meroni;M. Nava Giulia;S. Sertic;P. Plevani;M. Muzi-Falconi;F. Lazzaro
2018

Abstract

Ribonucleotides (rNTPs) are incorporated into genomic DNA at a relatively high frequency during replication. They have beneficial effects but, if not removed from the chromosomes, increase genomic instability. Here, we describe a fast method to easily estimate the amounts of embedded ribonucleotides into the genome. The protocol described is performed in Saccharomyces cerevisiae and allows us to quantify altered levels of rNMPs due to different mutations in the replicative polymerase ε. However, this protocol can be easily applied to cells derived from any organism.
DNA polymerase; DNA repair; DNA replication; Genome stability; Genomic rNMPs; RNase H; Ribonucleotides incorporation
Settore BIO/11 - Biologia Molecolare
Settore BIO/18 - Genetica
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/551603
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