000			02172nam a2200241 4500
001			393968
090			_a393968
100			_a20160531d2014 y0pory50 ba
101			_aeng
200			_aMicrochip analyzer for forensic short tandem repeat typing of single cells _bRecurso eletrónico _efinal technical report _fTao Geng, Richard A. Mathies
210			_a[Rockville, MD] _cNational Criminal Justice Reference Service (NCJRS) _d2014
215			_a1 CD-ROM _d12 cm
300			_aResearch report submitted to the U.S. Department of Justice, Award Number: 2009-DN-BX-K180. Ficheiro de 3,15 MB em formato PDF (39 p.)
330			_aShort tandem repeat (STR) typing at the single-cell level is a promising tool for human forensicidentification when the biological evidence materials are comprised of mixtures of cells from multiple individuals at relatively low concentrations. Here we describe a novel single-cell STR typing method with high sensitivity, fidelity and throughput that combines microfluidic droplet generation with single-cell multiplex emulsion PCR. Individual cells are separately isolated within microdroplets that subsequently function as miniaturized reactors for PCR amplification, producing high quality STR profiles from single cells at high throughput. CONTENTS: 1. Introduction. 2. Methods. 2.1 Microfluidic device fabrication. 2.2 Preparation of primer-functionalized beads. 2.3 Cell culture and sample preparation. 2.4 Microdroplet generation. 2.5 Cell lysis and DNA purification. 2.6 Emulsion PCR. 2.7 Bead recovery. 2.8 Secondary PCR and fragment sizing analysis. 3. Results. 3.1 Microbead-based solid-phase PCR in bulk solutions. 3.2 Agarose microdroplet-based emulsion PCR. 3.3 Single-molecule PCR in microdroplets. 3.4 Single-cell STR typing in microdroplets. 4. Conclusions. 5. References. 6. Dissemination of research findings.
606			_aIDENTIFICAÇÃO DE PESSOAS _9343105
606			_aQUÍMICA FORENSE _9343048
606			_aANÁLISE LABORATORIAL _9326721
700			_aGENG _bTao _9358134
701			_aMATHIES _bRichard A. _9358135
955			_xOferta