Design and operation of a portable scanner for high performance microchip capillary array electrophoresis

• NCJ Number: 307051
• Author(s): James R. Scherer; Peng Liu; Richard A. Mathies
• Date Published: 2010
• Grant Number(s): 2009-DN-BX-K180; 2007-DN-BX-K142
• Agencies: NIJ-Sponsored
• Publication Type: Research (Applied/Empirical)
• Award: Microchip Analyzer for Forensic Short Tandem Repeat Typing of Single Cells; Microfabricated Capillary Array Electrophoresis Genetic Analysis for Forensic Short Tandem Repeat DNA Profiling

Annotation

This article describes the development of a compact, laser-induced fluorescence detection scanner as a platform for electrophoretic detection and control of high-throughput, integrated microfluidic devices for genetic and other analyses, which can be used in high-performance forensic human identification.

Abstract

The authors have developed a compact, laser-induced fluorescence detection scanner, the multichannel capillary array electrophoresis portable scanner (McCAEPs) as a platform for electrophoretic detection and control of high-throughput, integrated microfluidic devices for genetic and other analyses. The instrument contains a confocal optical system with a rotary objective for detecting four different fluorescence signals, a pneumatic system consisting of two pressure/vacuum pumps and 28 individual addressable solenoid valves for control of on-chip microvalves and micropumps, four Polymerase Chain Reaction (PCR) temperature control systems, and four high voltage power supplies for electrophoresis. The detection limit of the instrument is ∼20 pM for on-chip capillary electrophoresis of fluorescein dyes. To demonstrate the system performance for forensic short tandem repeat (STR) analysis, two experiments were conducted: (i) electrophoretic separation and detection of STR samples on a 96-lane microfabricated capillary array electrophoresis microchip. Fully resolved PowerPlex® 16 STR profiles amplified from 1 ng of 9947A female standard DNA were successfully obtained; (ii) nine-plex STR amplification, sample injection, separation, and fluorescence detection of 100-copy 9948 male standard DNA in a single integrated PCR- capillary electrophoresis microchip. These results demonstrate that the McCAEPs can be used as a versatile control and detection instrument that operates integrated microfluidic devices for high-performance forensic human identification. (Publisher Abstract Provided)