biostudies-literatureKincaid KNIGMS NIH HHSe109https://www.ebi.ac.uk/biostudies/studies/S-EPMC1636374biostudies-literatureUnknown34(16)There has been a long-standing interest in the discovery of unnatural nucleotides that can be incorporated into DNA by polymerases. However, it is difficult to predict which nucleotide analogs will prove to have biological relevance. Therefore, we have developed a new screening method to identify novel substrates for DNA polymerases. This technique uses the polymerase itself to select a dNTP from a pool of potential substrates via incorporation onto a short oligonucleotide. The unnatural nucleotide(s) is then identified by high-resolution mass spectrometry. By using a DNA polymerase as a selection tool, only the biologically relevant members of a small nucleotide library can be quickly determined. We have demonstrated that this method can be used to discover unnatural base pairs in DNA with a detection threshold of < or =10% incorporation.Nucleic acids researchA mass spectrometry-based approach for identifying novel DNA polymerase substrates from a pool of dNTP analogues.PMC1636374R01 GM054194GM54194Kuchta RDKincaid KfalseA mass spectrometry-based approach for identifying novel DNA polymerase substrates from a pool of dNTP analogues.There has been a long-standing interest in the discovery of unnatural nucleotides that can be incorporated into DNA by polymerases. However, it is difficult to predict which nucleotide analogs will prove to have biological relevance. Therefore, we have developed a new screening method to identify novel substrates for DNA polymerases. This technique uses the polymerase itself to select a dNTP from a pool of potential substrates via incorporation onto a short oligonucleotide. The unnatural nucleotide(s) is then identified by high-resolution mass spectrometry. By using a DNA polymerase as a selection tool, only the biologically relevant members of a small nucleotide library can be quickly determined. We have demonstrated that this method can be used to discover unnatural base pairs in DNA with a detection threshold of < or =10% incorporation.2006-01-01T00:00:00Z20062024-02-14T23:23:52.864Z2019-03-27T01:46:14ZS-EPMC16363741694594910.1093/nar/gkl632