We propose to develop a low-cost 10-Gb Pyrosequencer for de novo DNA sequencing that would enable any lab to perform high-throughput genome analyses. The platform implements automated sample preparation scheme combined with massive Pyrosequencing, which will potentially enable mammalian genome sequencing in a single run. A sensitive CMOS image sensor has been designed and fabricated specialized for Pyrosequencing chemistry, which is integrated with fluidic platform. Each well in the fluidic platform has a pixel on CMOS for detection of light signal generated from Pyrosequencing. The integrated chip will have a 2-megapixel CMOS, each pixel located on a single well enabling 2 million sequencing on each chip. Sixteen of such chips are set up on a board to develop a 32-million wells Pyrosequencer which potentially enable sequencing of more than 10 gigabase of genomic DNA. The raw data are immediately collected and assembled with our developed algorithms. We envision that the genome sequencing of mammalian genome to be reduced below $100,000 level for large-scale genome sequencing projects. PROJECT

Public Health Relevance

This proposal aims to develop a new technology enabling rapid and high throughput DNA sequencing at any lab or clinic. This technology would have direct impact on rapid and accurate diagnosis, identification of new drug target, rapid environmental monitoring and would enhance our understanding about biological systems.

Agency
National Institute of Health (NIH)
Institute
National Human Genome Research Institute (NHGRI)
Type
Research Project (R01)
Project #
5R01HG003571-06
Application #
7676224
Study Section
Special Emphasis Panel (ZHG1-HGR-N (M1))
Program Officer
Schloss, Jeffery
Project Start
2004-09-30
Project End
2011-05-31
Budget Start
2009-06-01
Budget End
2010-05-31
Support Year
6
Fiscal Year
2009
Total Cost
$1,684,296
Indirect Cost
Name
Illumina, Inc.
Department
Type
DUNS #
033305264
City
San Diego
State
CA
Country
United States
Zip Code
92122
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Sundquist, Andreas; Bigdeli, Saharnaz; Jalili, Roxana et al. (2007) Bacterial flora-typing with targeted, chip-based Pyrosequencing. BMC Microbiol 7:108
Parameswaran, Poornima; Jalili, Roxana; Tao, Li et al. (2007) A pyrosequencing-tailored nucleotide barcode design unveils opportunities for large-scale sample multiplexing. Nucleic Acids Res 35:e130
Sundquist, Andreas; Ronaghi, Mostafa; Tang, Haixu et al. (2007) Whole-genome sequencing and assembly with high-throughput, short-read technologies. PLoS One 2:e484
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Agah, Ali; Aghajan, Mariam; Mashayekhi, Foad et al. (2004) A multi-enzyme model for Pyrosequencing. Nucleic Acids Res 32:e166