Droplet enhanced microfluidic-based DNA purification from bacterial lysates via phenol extraction |
| |
Authors: | Mercedes C Morales Jeffrey D Zahn |
| |
Affiliation: | (1) BioMEMS Laboratory, Department of Biomedical Engineering, Rutgers, The State University of New Jersey, Room 370, 599 Taylor Road, Piscataway, NJ 08854, USA;(2) BioMEMS Laboratory, Department of Biomedical Engineering, Rutgers, The State University of New Jersey, Room 311, 599 Taylor Road, Piscataway, NJ 08854, USA |
| |
Abstract: | Microfluidic platforms have been developed to demonstrate DNA purification via liquid extraction techniques at the microscale
using an aqueous phase containing either protein, DNA, or a complex cell lysate and an immiscible receiving organic (phenol)
phase. Initially, a serpentine device was used to investigate protein partitioning between the aqueous and organic phase,
and DNA purification when both protein and DNA were mixed in the aqueous phase and infused conjunctly with the phenol phase.
This two-phase system was studied using both stratified and droplet-based flow conditions. The droplet-based flow resulted
in a significant improvement of protein partitioning from the aqueous phase into the organic phase due to the convective flow
recirculation inside each droplet improving material transport to the organic–aqueous interface. A second device was designed
and fabricated to specifically extract plasmid DNA from bacterial lysates using only droplet-based flows. The plasmid recovery
using the microdevice was high (>92%) and comparable to the recovery achieved using commercial DNA purification kits and standard
macroscale phenol extraction. This study presents the initial steps toward the miniaturization of an efficient on-chip DNA
sample preparation using phenol extraction which could be integrated with post-extraction DNA manipulations for integrated
genomic analysis modules. |
| |
Keywords: | |
本文献已被 SpringerLink 等数据库收录! |
|