A sensitive assay for dNTPs based on long synthetic oligonucleotides, EvaGreen dye and inhibitor-resistant high-fidelity DNA polymerase
University of Helsinki · Folkhälsans Forskningscentrum · +4 more institutions
Abstract
Deoxyribonucleoside triphosphates (dNTPs) are vital for the biosynthesis and repair of DNA. Their cellular concentration peaks during the S phase of the cell cycle. In non-proliferating cells, dNTP concentrations are low, making their reliable quantification from tissue samples of heterogeneous cellular composition challenging. Partly because of this, the current knowledge related to the regulation of and disturbances in cellular dNTP concentrations derive mostly from cell culture experiments with little corroboration at the tissue or organismal level. Here, we fill the methodological gap by presenting a simple non-radioactive microplate assay for the quantification of dNTPs with a minimum requirement of 4-12…
Citation impact
- FWCI
- 41.84
- Percentile
- 100%
- References
- 45
Authors
5- JPJanne PurhonenCorresponding
University of Helsinki, Folkhälsans Forskningscentrum
- RBRishi Banerjee
University of Helsinki, Folkhälsans Forskningscentrum
- AEAllison E. McDonald
Wilfrid Laurier University
- VFVineta Fellman
University of Helsinki, Lund University, Helsinki University Hospital, Helsinki Children's Hospital, Folkhälsans Forskningscentrum
- JKJukka Kallijärvi
University of Helsinki, Folkhälsans Forskningscentrum
Topics & keywords
- Biology
- DNA polymerase
- Oligonucleotide
- DNA
- DNA synthesis
- DNA replication
- Molecular biology
- Taq polymerase
- Reduced inequalities