The design and in vitro synthesis of genes or long DNA molecules is an important and powerful tool in biomedical research, synthetic biology and protein engineering. While the actual synthesis and assembly reactions are highly amenable to automation and parallel processing, the major bottlenecks for making in vitro DNA synthesis a routine, high throughput and high speed process are the verification and recovery of the full length and error free synthetic DNA molecules which rely on laborious E. coli cloning and Sanger sequencing technologies. We propose the development of a bench top microfluidic system that has the ability to perform DNA synthesis, sequence verification and recovery of the desired full length DNA molecules. The system incorporates the innovative and flexible digital microfluidic platform developed by Advanced Liquid Logic, Inc. (ALL) with an efficient gene assembly protocol developed by the J. Craig Venter Institute (JCVI). The resulting system will employ pyrosequencing chemistry on a digital microfluidic cartridge to enable sequence verification and recovery of full length, error- free DNA molecules bypassing the laborious cloning and capillary based sequencing processes. The system will also enable direct coupling of the microchip-based oligonucleotide synthesis platform to the downstream assembly, sequencing and recovery process, further reducing the costs and improving the overall efficiency of gene and genome synthesis.

Public Health Relevance

(provided by applicant): The design and in vitro synthesis of genes or long DNA molecules is an important and powerful tool in biomedical research, synthetic biology and protein engineering. We propose the development of a bench top microfluidic system that has the ability to perform DNA synthesis, sequence verification and recovery of the desired full length DNA molecules.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21GM104694-01
Application #
8352845
Study Section
Special Emphasis Panel (ZRR1-BT-7 (01))
Program Officer
Friedman, Fred K
Project Start
2012-09-01
Project End
2015-08-31
Budget Start
2012-09-01
Budget End
2013-08-31
Support Year
1
Fiscal Year
2012
Total Cost
$248,770
Indirect Cost
$59,498
Name
J. Craig Venter Institute, Inc.
Department
Type
DUNS #
076364392
City
Rockville
State
MD
Country
United States
Zip Code
20850