ArticleAMB Express2022
DAR-PCR: a new tool for efficient retrieval of unknown flanking genomic DNA.
Article in AMB Express, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 18 citations in OpenAlex.
- Article
- Bridging PCR-Based Genome-Walking Protocol.Bio-protocol · 2025Article
- Center Degenerated Walking-Primer PCR: A Novel and Universal Genome-Walking Method.Current issues in molecular biology · 2025Article
- Uracil walking primer PCR: An accurate and efficient genome-walking tool.Journal, genetic engineering & biotechnology · 2025Article
- Protocol to Mine Unknown Flanking DNA Using PER-PCR for Genome Walking.Bio-protocol · 2025Article
- Protocol to Identify Unknown Flanking DNA Using Partially Overlapping Primer-based PCR for Genome Walking.Bio-protocol · 2025Article
- Protocol to Retrieve Unknown Flanking DNA Using Fork PCR for Genome Walking.Bio-protocol · 2025Article
- Thiamine-Responsive Megaloblastic Anemia Syndrome Mimicking Myelodysplastic Neoplasm.Acta haematologica · 2025Article
- Primer extension refractory PCR: an efficient and reliable genome walking method.Molecular genetics and genomics : MGG · 2024Article
- Sodium-Ion-Free Fermentative Production of GABA withMetabolites · 2023Article
- Semi-Site-Specific Primer PCR: A Simple but Reliable Genome-Walking Tool.Current issues in molecular biology · 2023Article
- Bridging PCR: An Efficient and Reliable Scheme Implemented for Genome-Walking.Current issues in molecular biology · 2023Article
- Fork PCR: a universal and efficient genome-walking tool.Frontiers in microbiology · 2023Article
- Fusion primer driven racket PCR: A novel tool for genome walking.Frontiers in genetics · 2022Article
Corrections and comments
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Authors and funding
8 authors at 3 institutions in 2 countries.
Funding
Abstract
Various PCR-based genome-walking methods have been developed to acquire unknown flanking DNA sequences. However, the specificity and efficacy levels, and the operational processes, of the available methods are unsatisfactory. This work proposes a novel walking approach, termed differential annealing-mediated racket PCR (DAR-PCR). The key to DAR-PCR is the use of primer-mediated intra-strand annealing (ISA). An ISA primer consists of a 5' root homologous to the known sequence and a heterologous 3' bud. In the single low-stringency cycle, the ISA primer anneals to a site on an unknown region and extends towards the sequence-specific primer (SSP) 1 site, thereby forming a target single-stranded DNA bound by the SSP1 complement and the ISA primer. In the subsequent more stringent cycles, its complementary strand is accumulated, owing to the differential annealing between the moderate-stringency ISA primer and the high-stringency SSP1. The accumulation of this strand provides an opportunity for ISA mediated by the ISA primer root. A loop-back extension subsequent to ISA occurs, creating a racket-like DNA with the known region positioned at both ends of the unknown sequence. This DNA is exponentially amplified during the secondary PCR driven by an SSP pair inner to SSP1. DAR-PCR was validated as an efficient walking method by determining unknown flanking sequences in Lactobacillus brevis and Oryza sativa.
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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.