BAC Library Production /Distribuion for 'Healthy people

DESCRIPTION: (provided by applicant) Cloning large DNA fragments using the
bacterial artificial chromosome (BAC) library system has revolutionized and
accelerated our ability to physically map, positionally clone and sequence
whole genomes. Due to the difficulty in cloning large DNA fragments, only a
handful of laboratories have been able to produce deep-coverage large insert
libraries on a routine basis. This bottleneck has resulted in a limited number
of BAC libraries that are available to the research community that can be used
for gene discovery, survey sequencing and comparative genome studies. To
increase the world?s BAC library construction capacity and reduce costs, the
NIH has posted a request for applications (RFA.HG-Ol-002) entitled "BAC Library
Production" which is the subject of this application.
The specific aims of our thee-year proposal are:
BAC Library Production: Develop BAC library construction capacity to construct
36 human genome size ( about3000 Mb) 10X genome equivalent BAC libraries. In
year 1, we will produce 6 libraries at $0.50/clone, in year 2, 12 libraries
will be produced at $0.375/clone and in year three 18 libraries will he
produced at $0.25/clone. The initial average insert size will hover between 130
- 150 kb and we will strive to increase our average insert sizes to 170 - 200
kb in subsequent years. Libraries, filters and clones will be distributed
through our CUGI BAC/EST Resource Center on a cost recovery basis
(www.genome.clemson.edu).
BAC Library Research and Development: We will systematically develop
technologies to optimize and streamline BAC library construction procedures in
order to reduce the cost of making libraries while maintaining quality and
insert size. Also, technologies will be developed to routinely achieve insert
sizes of 160kb or greater. In addition, a robust tool kit will be established
to produce high-quality megabase-size DNA from animals. This will include the
development of a "BIG DNA To-Go" kit to produce high-quality megabase-size DNA
on location where the lab needing a library will provide the tissue(s) required
to make the library.
Bioinformatics Support: A laboratory information management/bar coding system
will be developed to aid in tracking library production and distribution and a
robust online BAC library ordering system and database will also be developed.