This is a new Phase I proposal seeking support to develop an new cloning vector for use in shotgun DNA sequencing strategies. It is argued that currently, shotgun DNA sequencing approaches, which dominate current high throughput efforts in the US, are in part limited by the requirement to produce enough templates to carry out the approximately 5-fold redundant sequencing of the human genome. The applicant proposes to develop a sequencing vector in which it may be possible to clone five DNA inserts simultaneously and to carry out bi-directional reads on each insert (i.e. 10 reads). This can reduce the number of template preparations required by up to 10 fold depending on exact strategy and the efficiency of cloning. It may be even possible to multiplex sequencing runs which could further reduce sequencing efforts downstream of the template preparation step.
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