Abstract
In this paper we describe a novel method for detecting many DNA fragments through efficient amplification by using an emulsion system based on “on-chip” PCR instead of conventional multiplex polymerase chain reaction (PCR). During the preparation of on-chip PCR, a set of primers were immobilized on a slide and other sets were in an emulsion system. Different emulsion phase primers and other related PCR components were dispersed in different droplets of the emulsion system, and then, due to the thermal instability of emulsion droplets, they would be released onto the surface of the slide after preheating in the first PCR step. To test the above method, we used plasma DNAs from pregnant women who was carrying a male fetus for gender identification. Four different Y chromosome DNA fragments were selected. Results showed that different DNA fragments could be simultaneously amplified with satisfactory results. It is suggested that a simple, convenient and inexpensive on-chip PCR method has been developed.
Highlights
Multiplex polymerase chain reaction (PCR), along with PCR, is one of the most commonly used techniques in molecular biology
Multiplex PCR, along with PCR, is one of the most commonly used techniques in molecular biology. It is an essential cost-saving technique for large scale genotyping with significant scientific, clinical, and commercial applications, including gene expression [1], whole-genome sequencing [2], facilitating the diagnosis of infectious diseases [3] and forensic analysis, including human identification and paternity testing [4]. It often serves as the first step in many genetic analysis methods and a number of analytical methods are available for the detection of PCR products
The decrease of interferences contributed to the solution phase primers in the droplets of emulsion system where all molecules required in PCR is separated before thermal cycle
Summary
Multiplex PCR, along with PCR, is one of the most commonly used techniques in molecular biology It is an essential cost-saving technique for large scale genotyping with significant scientific, clinical, and commercial applications, including gene expression [1], whole-genome sequencing [2], facilitating the diagnosis of infectious diseases [3] and forensic analysis, including human identification and paternity testing [4]. It often serves as the first step in many genetic analysis methods and a number of analytical methods are available for the detection of PCR products. We have attempted to develop an emulsion based on-chip polymerase chain (EC-PCR) reaction in this study
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