ArticleFrontiers in genetics2022
Fusion primer driven racket PCR: A novel tool for genome walking.
Article in Frontiers in genetics, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
- Bridging PCR-Based Genome-Walking Protocol.Bio-protocol · 2025Article
- Article
- NBiochemical genetics · 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
- 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
- Bridging PCR: An Efficient and Reliable Scheme Implemented for Genome-Walking.Current issues in molecular biology · 2023Article
- Semi-Site-Specific Primer PCR: A Simple but Reliable Genome-Walking Tool.Current issues in molecular biology · 2023Article
- Fork PCR: a universal and efficient genome-walking tool.Frontiers in microbiology · 2023Article
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The limitations of the current genome-walking strategies include strong background and cumbersome experimental processes. Herein, we report a genome-walking method, fusion primer-driven racket PCR (FPR-PCR), for the reliable retrieval of unknown flanking DNA sequences. Four sequence-specific primers (SSP1, SSP2, SSP3, and SSP4) were sequentially selected from known DNA (5'→3') to perform FPR-PCR. SSP3 is the fragment that mediates intra-strand annealing (FISA). The FISA fragment is attached to the 5' end of SSP1, generating a fusion primer. FPR-PCR comprises two rounds of amplification reactions. The single-fusion primary FPR-PCR begins with the selective synthesis of the target first strand, then allows the primer to partially anneal to some place(s) on the unknown region of this strand, producing the target second strand. Afterward, a new first strand is synthesized using the second strand as the template. The 3' end of this new first strand undergoes intra-strand annealing to the FISA site, followed by the formation of a racket-like DNA by a loop-back extension. This racket-like DNA is exponentially amplified in the secondary FPR-PCR performed using SSP2 and SSP4. We validated this FPR-PCR method by identifying the unknown flanks of
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