ArticleFrontiers in bioengineering and biotechnology2022
Wristwatch PCR: A Versatile and Efficient Genome Walking Strategy.
Article in Frontiers in bioengineering and biotechnology, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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17 citing papers in PubMed.
- Development of Approaches for Transgene Expression in the Pathogenic Free-Living AmoebaPathogens (Basel, Switzerland) · 2025Article
- Peas in Rouge: Tyrosine Supplementation Enhances RUBY Reporter Visibility inPlants (Basel, Switzerland) · 2025Article
- Bridging PCR-Based Genome-Walking Protocol.Bio-protocol · 2025Article
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- Uracil walking primer PCR: An accurate and efficient genome-walking tool.Journal, genetic engineering & biotechnology · 2025Article
- Transgene Mapping in Animals: What to Choose?International journal of molecular sciences · 2025Review
- 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
- Hybrid substrate-based pH autobuffering GABA fermentation by Levilactobacillus brevis CD0817.Bioprocess and biosystems engineering · 2024Article
- 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
- Article
- Fusion primer driven racket PCR: A novel tool for genome walking.Frontiers in genetics · 2022Article
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Authors and funding
9 authors.
Funding
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Abstract
Genome walking is a method used to retrieve unknown flanking DNA. Here, we reported wristwatch (WW) PCR, an efficient genome walking technique mediated by WW primers (WWPs). WWPs feature 5'- and 3'-overlap and a heterologous interval. Therefore, a wristwatch-like structure can be formed between WWPs under relatively low temperatures. Each WW-PCR set is composed of three nested (primary, secondary, and tertiary) PCRs individually performed by three WWPs. The WWP is arbitrarily annealed somewhere on the genome in the one low-stringency cycle of the primary PCR, or directionally to the previous WWP site in one reduced-stringency cycle of the secondary/tertiary PCR, producing a pool of single-stranded DNAs (ssDNAs). A target ssDNA incorporates a gene-specific primer (GSP) complementary at the 3'-end and the WWP at the 5'-end and thus can be exponentially amplified in the next high-stringency cycles. Nevertheless, a non-target ssDNA cannot be amplified as it lacks a perfect binding site for any primers. The practicability of the WW-PCR was validated by successfully accessing unknown regions flanking
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