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An Enhanced Method For Sequence Walking And Paralog Mining: Topo® Vector-Ligation Pcr, Benjamin B. Orcheski, Thomas M. Davis

Biological Sciences

Background

Although technological advances allow for the economical acquisition of whole genome sequences, many organisms' genomes remain unsequenced, and fully sequenced genomes may contain gaps. Researchers reliant upon partial genomic or heterologous sequence information require methods for obtaining unknown sequences from loci of interest. Various PCR based techniques are available for sequence walking - i.e., the acquisition of unknown DNA sequence adjacent to known sequence. Many such methods require rigid, elaborate protocols and/or impose narrowly confined options in the choice of restriction enzymes for necessary genomic digests. We describe a new method, TOPO^® Vector-Ligation PCR (or TVL-PCR) that innovatively …