Abstract:

PCR-mediated approaches for genomic walking (or chromosome walking) are routinely used for cloning flanking sequences of the known DNA regions, which is an important technology for genomic research. According to the principles, they can be categorized into two types of ligation-dependence and ligation-independence. At present, numerous approaches of the later, each with distinct features, have been rapidly developed. Nevertheless, regarding the binding patterns of walking primers on DNA templates, they can be overall grouped into the degenerate primer-based or specific sequence-dependent subsets. Following this viewpoint, this paper summarized the major or representative techniques and the recent developments of ligation-independent, PCR-mediated genomic walking, so as to provide a methodological reference for the relevant researches.

Key words: ligation-independence, PCR, genomic walking, chromosome walking

摘要：

基于PCR的基因组步移(或染色体步移)技术常用于克隆已知DNA片段的旁侧序列，是一项重要的的基因组学研究技术。根据原理可将其分为依赖酶连介导和不需酶连介导两大类。目前后者发展迅速，方法种类较多而且各具特色，但根据步移引物与模板DNA的结合特点，可将其统分为基于简并引物的或依赖特定位点的基因组步移技术。综述了当前主要或典型的非酶连PCR介导的基因组步移方法以及最新进展，以期为相关研究提供方法借鉴。

关键词: 非酶连;PCR;基因组步移;染色体步移