High resolution DNA separations using microchip electrophoresis

Abstract

Planar microfluidic devices have emerged as effective tools for the electrophoretic separation of a variety of different DNA inputs. The advancement of this miniaturized platform was inspired initially by demands placed on electrophoretic performance metrics by the human genome project and has provided a viable alternative to slab gel and even capillary formats due to its ability to offer high resolution separations of nucleic acid materials in a fraction of the time associated with its predecessors, consumption of substantially less sample and reagents while maintaining the ability to perform many separations in parallel for realizing ultra-high throughputs. Another compelling advantage of this separation platform is that it offers the potential for integrating front-end sample preprocessing steps onto the separation device eliminating the need for manual sample handling. This review aims to compile a recent survey of various electrophoretic separations using either glass or polymer-based microchips in the areas of genotyping and DNA sequencing as well as those involving the growing field of DNA-based forensics.

Bioanalytical separations using electric field gradient techniques

“…The PFSG studies were often compared with slab gel and/or ME. In general, ME gives higher throughput (over 650 000 bases per 8 h versus under 70 000) and better resolution than slab gels, with a faster analysis time (tens of minutes versus hours) [129][130][131]. The ME-PFSG method can be used in conjunction with other techniques to improve DNA analysis.…”

Restriction Fragment Length Polymorphism Analysis of PCR-Amplified Fragments (PCR-RFLP) and Gel Electrophoresis - Valuable Tool for Genotyping and Genetic Fingerprinting

“…Recently, capillary electrophoresis and microchannel electrophoresis have become increasingly popular. They offer higher resolving power and throughput than conventional slab gels (Stellwagen et al, 2009;Sinville & Soper, 2007).…”

“…Electrophoresis in microchips and microfluidic systems provides significant lower separation time and higher throughput compared to other types of electrophoretic systems (Sinville & Soper, 2007). Another advantage of the microchip format is that electrophoresis can be integrated with the other steps of a PCR-RFLP procedure.…”

“…Microchips can also be produced using a variety of polymers (Sinville & Soper, 2007). Microchips are fabricated using techniques from the semiconductor and plastics industries (Fiorini & Chiu, 2005).…”

DNA Purification on Microfluidic Devices with a Focus on Large Volume, Forensic Biological Samples

“…24 Numerous processes have been demonstrated on microfluidics including, but not limited, nucleic acid extraction 9,[25][26][27][28][29][30][31][32] , PCR [33][34][35][36] , and separation/detection of PCR and sequencing products 10,19,[37][38][39][40][41][42] . The integration of these processes on a single device provides a closed system which eliminates the number of sample transfer steps involved and therefore decreases the points for entrance of contaminants or DNases.…”