Sorted libraries of error-free long oligonucleotides

Award Information
Agency:
Department of Health and Human Services
Branch
n/a
Amount:
$307,411.00
Award Year:
2012
Program:
SBIR
Phase:
Phase I
Contract:
1R44GM105192-01
Award Id:
n/a
Agency Tracking Number:
R44GM105192
Solicitation Year:
2012
Solicitation Topic Code:
NIGMS
Solicitation Number:
PA12-088
Small Business Information
5692 Plymouth rd, ANN ARBOR, MI, 48105-
Hubzone Owned:
N
Minority Owned:
N
Woman Owned:
N
Duns:
611643813
Principal Investigator:
JEAN-MARIE ROUILLARD
(734) 763-4722
jmr@biodiscovery-llc.com
Business Contact:
JEAN ROUILLARD
(734) 998-0751
jmr@biodiscovery-llc.com
Research Institution:
Stub




Abstract
DESCRIPTION: Synthetic Biology is full of promises ranging from discovery and production of new drugs, targeted therapies modifying organisms such as yeast or E. coli or creating totally new ones, higher crop yields, CO2 sequestration to bio-energy projects. However, these goals have been difficult to reach primarily due to the difficulty of De Novo gene synthesis and synthesis of new control and regulatory metabolic pathways all the way to synthetic chromosomes from chemically synthesized DNA. Just like any reversible chemical reaction DNA fragment synthesis by sequentially adding the monomers (A,C,G,T) gives less than 100% yield at each step( typical stepwise yields are ~97-99%) as a result the longer the length of DNA to be synthesized the smaller is thefraction of pure product at the end. Elimination of the errors take significant time and money. The goal of this project is to provide error free long oligonucleotides (100 to 300 mers) at a cost lower than the impure DNA fragments of today to unlock thepotential of synthetic biology. We are proposing to make libraries of clonally amplified and sequence verified long oligonucleotides. Our long term objectives are to implement a commercial service of affordable custom synthesis of sequence-verified longoligonucleotide libraries. During Phase I, we will 1) to demonstrate that single oligonucleotide molecules can be clonally amplified and sequence verified, 2) to demonstrate that beads bearing clonal amplifications can be captured on a microarray bearingsequence-specific probes and 3) to demonstrate that sequence verified oligonucleotides enable gene assembly with ten fold reduced error rate. During Phase II, we will 1) to develop a protocol/device to normalize the number of each oligonucleotide surveyedfrom a library, 2) to establish a standard operating procedure for the production of large libraries of sequence-verified oligonucleotides and 3) to determine the maximum oligonucleotide length that could be offered as a commercial product. PUBLIC HEALTH RELEVANCE: The unprecedented availability of affordable custom libraries of sequence verified long oligonucleotides will enable faster, easier and cheaper gene and large DNA fragment assembly with major applications in Synthetic Biology. This technology will undoubtedly bolster the discovery of new cellular mechanisms, molecular therapeutic tools, or even drugs, ultimately benefiting the society.

* information listed above is at the time of submission.

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