ArticleNucleic acids research2011
Efficient preparation of internally modified single-molecule constructs using nicking enzymes.
Article in Nucleic acids research, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed, 45 citations in OpenAlex.
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- De novo fabrication of custom-sequence plasmids for the synthesis of long DNA constructs with extrahelical features.Biophysical journal · 2024Article
- Dynamic interplay between target search and recognition for a Type I CRISPR-Cas system.Nature communications · 2023Article
- Efficient golden gate assembly of DNA constructs for single molecule force spectroscopy and imaging.Nucleic acids research · 2022Article
- Human HELB is a processive motor protein that catalyzes RPA clearance from single-stranded DNA.Proceedings of the National Academy of Sciences of the United States of America · 2022Article
- Distinct RPA domains promote recruitment and the helicase-nuclease activities of Dna2.Nature communications · 2021Article
- High-yield purification of exceptional-quality, single-molecule DNA substrates.Journal of biological methods · 2021Article
- Chemical roadblocking of DNA transcription for nascent RNA display.The Journal of biological chemistry · 2020Article
- A molecular device: A DNA molecular lock driven by the nicking enzymes.Computational and structural biotechnology journal · 2020Article
- High-yield fabrication of DNA and RNA constructs for single molecule force and torque spectroscopy experiments.Nucleic acids research · 2019Article
- Competing interaction partners modulate the activity of Sgs1 helicase during DNA end resection.The EMBO journal · 2019Article
- Single molecule glycosylase studies with engineered 8-oxoguanine DNA damage sites show functional defects of a MUTYH polyposis variant.Nucleic acids research · 2019Article
- Multiple RPAs make WRN syndrome protein a superhelicase.Nucleic acids research · 2018Article
- Efficient modification of λ-DNA substrates for single-molecule studies.Scientific reports · 2017Article
- Fluorescently labeled circular DNA molecules for DNA topology and topoisomerases.Scientific reports · 2016Article
- Unwinding forward and sliding back: an intermittent unwinding mode of the BLM helicase.Nucleic acids research · 2015Article
- Locked nucleic acid oligomers as handles for single molecule manipulation.Nucleic acids research · 2014Article
- Nuclease activity of Saccharomyces cerevisiae Dna2 inhibits its potent DNA helicase activity.Proceedings of the National Academy of Sciences of the United States of America · 2013Article
Corrections and comments
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Authors and funding
7 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Investigations of enzymes involved in DNA metabolism have strongly benefited from the establishment of single molecule techniques. These experiments frequently require elaborate DNA substrates, which carry chemical labels or nucleic acid tertiary structures. Preparing such constructs often represents a technical challenge: long modified DNA molecules are usually produced via multi-step processes, involving low efficiency intermolecular ligations of several fragments. Here, we show how long stretches of DNA (>50 bp) can be modified using nicking enzymes to produce complex DNA constructs. Multiple different chemical and structural modifications can be placed internally along DNA, in a specific and precise manner. Furthermore, the nicks created can be resealed efficiently yielding intact molecules, whose mechanical properties are preserved. Additionally, the same strategy is applied to obtain long single-strand overhangs subsequently used for efficient ligation of ss- to dsDNA molecules. This technique offers promise for a wide range of applications, in particular single-molecule experiments, where frequently multiple internal DNA modifications are required.
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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.