ArticleScientific reports2013
Randomly broken fragment PCR with 5' end-directed adaptor for genome walking.
Article in Scientific reports, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.
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Who cites it
4 citing papers in PubMed.
- Advances in CRISPR-enabled genome-wide screens in yeast.FEMS yeast research · 2025Review
- Simple innovative adaptor to improve genome walking with convenient PCR.Journal, genetic engineering & biotechnology · 2020Article
- Cyclic Digestion and Ligation-Mediated PCR Used for Flanking Sequence Walking.Scientific reports · 2020Article
- ALF: a strategy for identification of unauthorized GMOs in complex mixtures by a GW-NGS method and dedicated bioinformatics analysis.Scientific reports · 2017Article
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7 authors.
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Abstract
Many genome walking methods based on polymerase chain reaction (PCR) are available, including those with and without restriction enzyme modification. Nevertheless, these methods suffer from low reproducibility, inefficiency, and non-specificity. Here, we present a traceable and efficient PCR strategy: randomly broken fragment PCR with 5' end-directed adaptor for genome walking. The genome is first fragmented randomly. After blunting ends, the fragments are ligated to the 5' end-directed adaptors. Semi-nested PCR is then performed. Thus, we can obtain an unknown sequence by cloning the fragments of interest, followed by sequencing. This method effectively bypasses the above-mentioned obstacles and offers the advances: 1) genome fragmentation without using restriction enzymes; 2) enhancement of primer specificity and the prevention of self-ligation between the adaptors by employing a 5' end-directed adaptor. All of the steps in this new method are straightforward, and the unknown sequence can be definitively obtained by merely applying the method once.
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