Work

Protocol to clone unknown flanking genomic region using SWPOP-PCR for genome walking

Year: 2025

Type: article

Abstract: Genome walking is a molecular technique for mining unknown flanking DNAs, significantly advancing fields related to biology. We here detail a genome-walking protocol for stepwise partially overlapping primer-based PCR (SWPOP-PCR). We specify the rules for designing walking primers and gene-specific primers. We also delineate the three-stage workflow of SWPOP-PCR for cloning target DNA, as well as the subsequent purification, sequencing, and analysis of target amplicon. For complete details on the use and execution of this protocol, please refer to Chang et al. 1

Source: STAR Protocols

DOI: 10.1016/j.xpro.2025.104220

Authors: Qingchun Tang, Kunpeng Chang, Mou Li, Haixing Li

Institution: Nanchang University

Language: English

Topic: Molecular Biology Techniques and Applications

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $1,550

Funders: Nanchang University, State Key Laboratory of Food Science and Technology, Major Discipline Academic and Technical Leaders Training Program of Jiangxi Province, State Key Laboratory of Food Science and Technology, Nanchang University