A strategy for the amplification, purification, and selection of M13 templates for large-scale DNA sequencing

摘要: A strategy addressing various aspects of generating and processing DNA sequencing templates is described. The designed for large-scale projects involves procedures the amplification, purification, selection M13 templates. amplification based on growth in liquid culture, purification a novel thermoextraction method which replaces phenol extraction ethanol precipitation, template hybridization gridded arrays membranes with chemiluminescent probes. All steps are currently carried out manually but use standardized formats (96-well format) throughout entire provides possible transition to partial or full automation future. has been developed tested within ongoing project class II region human major histocompatibility complex it should be equally applicable any project.