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A new approach using multiplex long accurate PCR and yeast artificial chromosomes for bacterial chromosome mapping
A Sorokin1, A Lapidus, V Capuano
1Laboratoire de Génétique Microbienne, Institut National de la Recherche Agronomique, Jouy en Josas, France. sorokine@biotec.jouy.inra.fr
Abstract:
An efficient approach for structural studies on bacterial chromosomes is presented. It is based on high-resolution PCR map construction by using a multiplex long accurate PCR (MLA PCR) protocol and a YAC clone carrying the region to be mapped as indicator. The high-resolution PCR map of the bacillus subtilis rrnB-dnaB region is presented as an example. Data are also presented on the use of DNA generated by LA PCR for sequencing; they are relevant to LA PCR induced mutations and justify the application of such mapping for sequencing long stretches of bacterial chromosomes.