ArticleNucleic acids research2016
A high-throughput assay for the comprehensive profiling of DNA ligase fidelity.
Article in Nucleic acids research, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers.
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Who cites it
30 citing papers in PubMed, 59 citations in OpenAlex.
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- Arrayed CRISPR libraries for the genome-wide activation, deletion and silencing of human protein-coding genes.Nature biomedical engineering · 2025Article
- Selection of Fusion-Site Overhang Sets for High-Fidelity and High-Complexity Golden Gate Assembly.Methods in molecular biology (Clifton, N.J.) · 2025Article
- GGAssembler: Precise and economical design and synthesis of combinatorial mutation libraries.Protein science : a publication of the Protein Society · 2024Article
- High-Fidelity RNA Copying via 2',3'-Cyclic Phosphate Ligation.Journal of the American Chemical Society · 2024Article
- Modular Assembly of Synthetic Secondary Chromosomes.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Small RNAs and Hfq capture unfolded RNA target sites during transcription.Molecular cell · 2023Article
- Quantitative analysis of miRNAs using SplintR ligase-mediated ligation of complementary-pairing probes enhanced by RNase H (SPLICER)-qPCR.Molecular therapy. Nucleic acids · 2023Article
- Profiling DNA Ligase Substrate Specificity with a Pacific Biosciences Single-Molecule Real-Time Sequencing Assay.Current protocols · 2023Article
- PlasmidMaker is a versatile, automated, and high throughput end-to-end platform for plasmid construction.Nature communications · 2022Article
- Mismatch discrimination and sequence bias during end-joining by DNA ligases.Nucleic acids research · 2022Article
- A comparison of mRNA sequencing (RNA-Seq) library preparation methods for transcriptome analysis.BMC genomics · 2022Article
- Capillary Electrophoresis-Based Functional Genomics Screening to Discover Novel Archaeal DNA Modifying Enzymes.Applied and environmental microbiology · 2022Article
- Structured sequences emerge from random pool when replicated by templated ligation.Proceedings of the National Academy of Sciences of the United States of America · 2021Article
- Cognate base-pair selectivity of hydrophobic unnatural bases in DNA ligation by T4 DNA ligase.Biopolymers · 2021Article
- Structural insight into DNA joining: from conserved mechanisms to diverse scaffolds.Nucleic acids research · 2020Review
- Riboswitch identification using Ligase-Assisted Selection for the Enrichment of Responsive Ribozymes (LigASERR).Synthetic biology (Oxford, England) · 2019Article
- Multiplex real-time PCR assay combined with rolling circle amplification (MPRP) using universal primers for non-invasive detection of tumor-related mutations.RSC advances · 2018Article
- A single-molecule sequencing assay for the comprehensive profiling of T4 DNA ligase fidelity and bias during DNA end-joining.Nucleic acids research · 2018Article
- Cyclization of secondarily structured oligonucleotides to single-stranded rings by usingRSC advances · 2018Article
Corrections and comments
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Authors and funding
8 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
DNA ligases have broad application in molecular biology, from traditional cloning methods to modern synthetic biology and molecular diagnostics protocols. Ligation-based detection of polynucleotide sequences can be achieved by the ligation of probe oligonucleotides when annealed to a complementary target sequence. In order to achieve a high sensitivity and low background, the ligase must efficiently join correctly base-paired substrates, while discriminating against the ligation of substrates containing even one mismatched base pair. In the current study, we report the use of capillary electrophoresis to rapidly generate mismatch fidelity profiles that interrogate all 256 possible base-pair combinations at a ligation junction in a single experiment. Rapid screening of ligase fidelity in a 96-well plate format has allowed the study of ligase fidelity in unprecedented depth. As an example of this new method, herein we report the ligation fidelity of Thermus thermophilus DNA ligase at a range of temperatures, buffer pH and monovalent cation strength. This screen allows the selection of reaction conditions that maximize fidelity without sacrificing activity, while generating a profile of specific mismatches that ligate detectably under each set of conditions.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.