Disposable on‐chip microfluidic system for buccal cell lysis, DNA purification, and polymerase chain reaction

Abstract

This paper reports the development of a disposable, integrated biochip for DNA sample preparation and PCR. The hybrid biochip (25 × 45 mm) is composed of a disposable PDMS layer with a microchannel chamber and reusable glass substrate integrated with a microheater and thermal microsensor. Lysis, purification, and PCR can be performed sequentially on this microfluidic device. Cell lysis is achieved by heat and purification is performed by mechanical filtration. Passive check valves are integrated to enable sample preparation and PCR in a fixed sequence. Reactor temperature is needed to lysis and PCR reaction is controlled within ±1°C by PID controller of LabVIEW software. Buccal epithelial cell lysis, DNA purification, and SY158 gene PCR amplification were successfully performed on this novel chip. Our experiments confirm that the entire process, except the off‐chip gel electrophoresis, requires only approximately 1 h for completion. This disposable microfluidic chip for sample preparation and PCR can be easily united with other technologies to realize a fully integrated DNA chip.

Microfluidic single-cell analysis—Toward integration and total on-chip analysis

“…65,66 Generally speaking, this approach is not compatible with continuous flow mode in dropletbased devices for instance, since it takes time to reach a desired temperature. In both microwell and valve formats, heat can be supplied to the device by built-in heating elements 67 [Fig. 4(b)]o r from external sources 29 [Fig.…”

DNA Purification by Solid Phase Extraction (SPE) Methods

“…Hence, researchers used magnetic based microfluidic system for purification spiked bacterial (E. coli and B. subtilis) DNA from human blood and amount of recovery DNA was evaluated with PCR study. In another study, disposable PDMS microchip was designed for multipurpose analysis [25]. Buccal cell lysis, DNA purification and selected gene PCR analysis were carried out on the same chip, then selected gene amplification result was compared with gel electrophoresis and interestingly all experimental results were completed in a short time nearly 1 hour.…”