DAR-PCR: a new tool for efficient retrieval of unknown flanking genomic DNA

Tianyi Sun, Mengya Jia, Lingqin Wang, Zhaoqin Li, Zhiyu Lin, Cheng Wei, Jinfeng Pei & Haixing Li
Abstract Various PCR-based genome-walking methods have been developed to acquire unknown flanking DNA sequences. However, the specificity and efficacy levels, and the operational processes, of the available methods are unsatisfactory. This work proposes a novel walking approach, termed differential annealing-mediated racket PCR (DAR-PCR). The key to DAR-PCR is the use of primer-mediated intra-strand annealing (ISA). An ISA primer consists of a 5’ root homologous to the known sequence and a heterologous 3’ bud. In the...
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