Microfluidic DNA amplification—A review

Abstract

The application of microfluidic devices for DNA amplification has recently been extensively studied. Here, we review the important development of microfluidic polymerase chain reaction (PCR) devices and discuss the underlying physical principles for the optimal design and operation of the device. In particular, we focus on continuous-flow microfluidic PCR on-chip, which can be readily implemented as an integrated function of a micro-total-analysis system. To overcome sample carryover contamination and surface adsorption associated with microfluidic PCR, microdroplet technology has recently been utilized to perform PCR in droplets, which can eliminate the synthesis of short chimeric products, shorten thermal-cycling time, and offers great potential for single DNA molecule and single-cell amplification. The work on chip-based PCR in droplets is highlighted.

Next-generation sequencing in the clinic: Promises and challenges

“…Second, multiplex amplification of target sequences using multiple primer sets in a single tube can result in an excessive amount of non-specific products. Several technologies have been developed to get around this problem, including microfluidic PCR [33,34], emulsion PCR [35,36], and microdroplet PCR [37]. These strategies enable many singleplex PCRs to be performed independently and simultaneously in a single reaction, where primer-pair crossinteractions and product-product hybridization are prevented.…”

Basic Technologies for Droplet Microfluidics

“…Other advantages of droplet-based [76] microfluidic system include precisely controllable reaction time inside a droplet by adjusting the length of channel and the flow rates of fluids. The availability of a wide range of technologies for flexible generation and manipulation of droplets has enabled the applications of droplet microfluidics in a wide variety of fields, from chemical reactions [9,79] and protein crystallization [10,11] to material synthesis [12,13,[80][81][82][83], single cell analysis [84][85][86][87][88][89][90][91], DNA amplification [16], protein engineering [62,92], and high-throughput screening technologies [17,93]. It is not possible to cover all application areas in this review.…”

“…Later on theoretical interests arose on the characterization and exploration of complex dynamic phenomena involved in droplet generation and trafficking within microfluidic channel networks [3,4]. Meanwhile, a wealth of technologies for improved droplet generation, intra-droplet content manipulation, and methods for controlled trafficking, all enabled a wide range of appealing applications [5][6][7][8] from chemical kinetics [9] and protein 70 S. Zeng et al crystallization [10,11] to material synthesis [12,13], single cell analysis [14,15], polymerase chain reaction (PCR) [16], protein engineering, and high throughput screening technologies [17]. This review will discuss the recent advances in technologies that enable droplets as microreactors with complete functions and the applications of droplets in chemistry and biology.…”

“…Applications of droplets in chemical reactions, particle synthesis, and single cell analysis will also be briefly reviewed. The readers are also addressed to some recent excellent reviews on various aspects of droplet microfluidics [5][6][7][8][9][14][15][16][17]. Studies from fluid mechanics perspectives, which focus on the understanding of the basic fluid dynamic phenomena involved in droplet or multiphase microfluidics in general, have been summarized in recent reviews [18][19][20][21] and are not detailed here.…”

A Rapid Magnetofluidic Micromixer Using Diluted Ferrofluid

“…Micromixers have a broad range of applications such as for reactors [2][3][4], lab on a chip for chemical engineering [5], enhancement of chemical selectivity [6], extraction processes [7], drug discovery [8], polymer synthesis [9], and DNA amplification [10].…”