ArticleCurrent issues in molecular biology2023
Semi-Site-Specific Primer PCR: A Simple but Reliable Genome-Walking Tool.
Article in Current issues in molecular biology, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 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
- Fork PCR: a universal and efficient genome-walking tool.Frontiers in microbiology · 2023Article
Corrections and comments
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Authors and funding
5 authors.
Funding
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Abstract
Genome-walking has been frequently applied to molecular biology and related areas. Herein, a simple but reliable genome-walking technique, termed semi-site-specific primer PCR (3SP-PCR), is presented. The key to 3SP-PCR is the use of a semi-site-specific primer in secondary PCR that partially overlaps its corresponding primary site-specific primer. A 3SP-PCR set comprises two rounds of nested amplification reactions. In each round of reaction, any primer is allowed to partially anneal to the DNA template once only in the single relaxed-stringency cycle, creating a pool of single-stranded DNAs. The target single-stranded DNA can be converted into a double-stranded molecule directed by the site-specific primer, and thus can be exponentially amplified by the subsequent high-stringency cycles. The non-target one cannot be converted into a double-strand due to the lack of a perfect binding site to any primer, and thus fails to be amplified. We validated the 3SP-PCR method by using it to probe the unknown DNA regions of rice hygromycin genes and
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