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Automated preparation of single-cells and nuclei from solid tissues

Award Information
Agency: Department of Health and Human Services
Branch: National Institutes of Health
Contract: 1R41HG010129-01
Agency Tracking Number: R41HG010129
Amount: $280,000.00
Phase: Phase I
Program: STTR
Solicitation Topic Code: 172
Solicitation Number: PA17-148
Solicitation Year: 2017
Award Year: 2018
Award Start Date (Proposal Award Date): 2018-06-13
Award End Date (Contract End Date): 2019-03-31
Small Business Information
Livermore, CA 94551-9409
United States
DUNS: 080361775
HUBZone Owned: No
Woman Owned: No
Socially and Economically Disadvantaged: No
Principal Investigator
 (925) 292-8243
Business Contact
Phone: (925) 413-5952
Research Institution
LA JOLLA, CA 92037-3498
United States

 Domestic Nonprofit Research Organization

Project Summary AbstractThis project designsdevelopsand automates a novel system to process solid tissue into
single cellssingle nucleiand high molecular weight DNA using disposable cartridges without
user interventionThe system will support downstream single cell RNA or DNA sequencingsingle nuclei RNA or DNA sequencingand long read sequencers and optical mappingThe
long term goal of the project is to enable researchers and clinicians with sample to answer
solutions for fully integrated sequencing of solid tissues including biopsy samplesIn the Phase I projectautomated workflows are developed and optimized in novelfully
integrated cartridge designsThe prototype system is first benchmarked against an Sbreadboard system and manual processes for single cell production from mouse tissuesCartridgesmechanical disruptionenzymologyand workflows are further developed and
optimized for mouse and rat tissues for fully integrated processing performanceA cartridge formg tissue specimens is designed and testedProcessing single cells into high molecular
weight DNA is tested using the magnetic separations in the cartridge and on other devicesThe automated preparation of nuclei will be tested and developed on prototype systemsSingle cell suspensions prepared on the prototype will be processed into nuclei by chemical lysis
in the cartridge and magnetic purification of nuclei with paramagnetic beads testedEnzymatic
formulations with gentler processing will be studiedWorkflows and cartridges to directly
process tissue into nuclei will be developed and testedThe single sample prototype systems will be tested externally at the J Craig Venter Institute
and the Broad InstitutePhase I testing will design the architecture of a multi sample preproduction prototype system with additional capabilities to be built and tested in Phase IIAn
optical module will be added to determine viability and titer on the system and potentially allow
the system to adjust dissociation protocolson the flyIntegrated magnetic processing will be
further developed to purify cell typesnuclei and other organellesand DNAThe project output will bea validated single sample tissue to cellnucleior high molecularweight DNA automated prototypeanda multiple sample design architected for Phase II
development Project NarrativeThis project develops a system to automatically process solid tissues into single cellsnucleiand high molecular weight DNAThe system addresses the pipeline gap of
preparing cells and nuclei from tissue for single cell next generation sequencingNGSThe system will allow the full power and scale of NGS to be applied to primary solid tissue
samples and will help standardize research and clinical applications of single cells and
nucleiMore broadlythe system will help create a fully integrated tissue to answer
sequencing system in the future

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

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