Introduction
Genome walking (GW) is a molecular procedure for the direct identification of nucleotide sequences from purified genomes, the only thing that we need to know is the nucleotide sequence from which to start. There are numerous methods for GW, and they can be classified into three main categories: (1). Restriction-based GW methods: it’s requiring a restriction digestion of genomic DNA and ligation of restriction fragments to DNA-cassettes; (2). Primer-based GW methods: the PCR amplification in this method are directly carried out using a variously designed combinatorial coupled to a sequence specific primer; (3). Extension-based GW methods: the extension of a sequence specific primer and subsequent 3’-tailing of the resulting single strand DNA (ssDNA) provide the substrate for the final PCR amplification (Fig. 1).
Fig. 1 Main GW strategies
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