We propose to develop efficient high-speed automated methods for the analysis of DNA sequence. Such methods are essential to the success of the Human Genome Initiative, in which the sequence of the human genome will be determined and used as a powerful tool for biological and medical research. Two aspects of sequence analysis will be addressed, the procedures in which DNA for sequencing is produced and fragmented, and the methods employed for separation and analysis. In the first area we will use the Polymerase Chain Reaction method in conjunction with support chemistry to produce pure sets of single-stranded DNA fragments for separation and detection. This chemistry will be automated using a combination of temperature ramping, filtration, and pipetting. We will also investigate the potential of phosphorothioate-based sequencing chemistry in automated sequencing. In the second area capillary electrophoretic separation of the DNA fragments will be investigated for its potential to dramatically increase speed and resolution in automated DNA sequencing.

Agency
National Institute of Health (NIH)
Institute
National Human Genome Research Institute (NHGRI)
Type
Research Project (R01)
Project #
5R01HG000321-03
Application #
3333416
Study Section
Special Emphasis Panel (SRC)
Project Start
1989-07-01
Project End
1994-06-30
Budget Start
1991-07-01
Budget End
1992-06-30
Support Year
3
Fiscal Year
1991
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Type
Schools of Arts and Sciences
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
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