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Improved Library Preparation Workflows for Next Generation Sequencing

Award Information
Agency: Department of Health and Human Services
Branch: National Institutes of Health
Contract: 1R43HG006820-01A1
Agency Tracking Number: R43HG006820
Amount: $132,803.00
Phase: Phase I
Program: SBIR
Solicitation Topic Code: NHGRI
Solicitation Number: PA12-088
Timeline
Solicitation Year: 2013
Award Year: 2013
Award Start Date (Proposal Award Date): N/A
Award End Date (Contract End Date): N/A
Small Business Information
9955 Mesa Rim Rd
San Diego, CA 92121-2911
United States
DUNS: 945720043
HUBZone Owned: No
Woman Owned: No
Socially and Economically Disadvantaged: No
Principal Investigator
 NATASHA PAUL
 (858) 546-0004
 npaul@trilinkbiotech.com
Business Contact
 RICHARD HOGREFE
Phone: (858) 546-0004
Email: rhogrefe@trilinkbiotech.com
Research Institution
 Stub
Abstract

DESCRIPTION (provided by applicant): Next generation sequencing (NGS) technologies are a rapidly developing area, with the ability to produce gigabases of sequencing data at reduced costs. While NGS is best recognized for genomic DNA sequencing (DNA-Seq),there is a significant body of work to expand the breadth of sample types being studied. RNA deep sequencing (RNA-Seq) is a well-developed segment of NGS experimentation, with a growing focus on small RNA deep sequencing (smRNA-Seq). One of the limitationsin the deep sequencing of the 20-30 nt small RNA is in the sample preparation workflow, which requires several purification steps. As with all RNA samples, NGS sample preparation involves the ligation of fixed sequences, called adapters, onto the 5 and 3ends of the starting RNA library. This ligation step can be plagued by the undesired joining of the adapter sequences to one another without a segment of the library in between, resulting in adapter dimer formation. Although affinity capture can be usedto remove adapter dimers in most library prep workflows, adapter-tagged small RNA libraries are too close in size to the adapter dimers for effective use of this approach. As result, a gel purification step is required, which can deplete low abundance sequences. To avoid making unintentional changes to the complexity of the library, it is advantageous to block adapter dimer formation at the ligation step. With this goal in mind, we propose a novel approach to RNA sample preparation for NGS that uses TriLink's expertise in nucleic acid chemistry to disfavor adapter-adapter ligation while allowing for efficient joining of adapter probes on the 5'and 3' ends of the library. PUBLIC HEALTH RELEVANCE PUBLIC HEALTH RELEVANCE: Next generation sequencing(NGS) is maturing into an essential technique for RNA and DNA sequence analysis. NGS is a major component of the genomics market which was valued at 5.8 billion in 2009. We propose the investigation of chemically-modified components to streamline NGS experimental workflows for small RNA deep sequencing.

* Information listed above is at the time of submission. *

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