ArticleNucleic acids research2003
T-linker-specific ligation PCR (T-linker PCR): an advanced PCR technique for chromosome walking or for isolation of tagged DNA ends.
Article in Nucleic acids research, 2003. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 34 papers.
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Who cites it
34 citing papers in PubMed.
- 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
- 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
- Identification of T-DNA structure and insertion site in transgenic crops using targeted capture sequencing.Frontiers in plant science · 2023Article
- Whole-genome sequencing identifies I-SceI-mediated transgene integration sites in Xenopus tropicalis snai2:eGFP line.G3 (Bethesda, Md.) · 2022Article
- Fusion primer driven racket PCR: A novel tool for genome walking.Frontiers in genetics · 2022Article
- Wristwatch PCR: A Versatile and Efficient Genome Walking Strategy.Frontiers in bioengineering and biotechnology · 2022Article
- Amplicon genome fishing (AGF): a rapid and efficient method for sequencing target cis-regulatory regions in nonmodel organisms.Molecular genetics and genomics : MGG · 2021Article
- Using Combined Methods of Genetic Mapping and Nanopore-Based Sequencing Technology to Analyze the Insertion Positions ofFrontiers in plant science · 2021Article
- Simple innovative adaptor to improve genome walking with convenient PCR.Journal, genetic engineering & biotechnology · 2020Article
- Mapping of transgenic alleles in soybean using a nanopore-based sequencing strategy.Journal of experimental botany · 2019Article
- A Simple Genome Walking Strategy to Isolate Unknown Genomic Regions Using Long Primer and RAPD Primer.Iranian journal of biotechnology · 2019Article
- Stepwise partially overlapping primer-based PCR for genome walking.AMB Express · 2018Article
- Linear and exponential TAIL-PCR: a method for efficient and quick amplification of flanking sequences adjacent to Tn5 transposon insertion sites.AMB Express · 2017Article
- The increased expression of follicle-stimulating hormone leads to a decrease of fecundity in transgenic Large White female pigs.Transgenic research · 2017Article
- Article
- Transcription coupled repair and biased insertion of human retrotransposon L1 in transcribed genes.Mobile DNA · 2017Article
- Identification of the Genomic Insertion Site of the Thyroid Peroxidase Promoter-Cre Recombinase Transgene Using a Novel, Efficient, Next-Generation DNA Sequencing Method.Thyroid : official journal of the American Thyroid Association · 2015Article
- Sequencing, identification and mapping of primed L1 elements (SIMPLE) reveals significant variation in full length L1 elements between individuals.BMC genomics · 2015Article
Corrections and comments
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Dozens of PCR-based methods are available for chromosome walking from a known sequence to an unknown region. These methods are of three types: inverse PCR, ligation-mediated PCR and randomly primed PCR. However, none of them has been generally applied for this purpose, because they are either difficult or inefficient. Here we describe a simple and efficient PCR strategy--T-linker-specific ligation PCR (T-linker PCR) for gene or chromosome walking. The strategy amplifies the template molecules in three steps. First, genomic DNA is digested with 3' overhang enzymes. Secondly, primed by a specific primer, a strand of the target molecule is replicated by Taq DNA polymerase and a single A tail is generated on the 3' unknown end of the target molecule, and then a 3' overhang-T linker (named T-linker) is specifically ligated onto the target. Thirdly, the target is amplified by two rounds of nested PCR with specific primers and T-linker primers. T-linker PCR significantly improves the existing PCR methods for walking because it uses specific T/A ligation instead of arbitrary ligation or random annealing. To show the feasibility and efficiency of T-linker PCR, we have exploited this method to identify vector DNA or T-DNA insertions in transgenic plants.
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