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可共聚寡聚微球的流动合成及其在单链DNA扩增中的应用。

On-Flow Synthesis of Co-Polymerizable Oligo-Microspheres and Application in ssDNA Amplification.

作者信息

Lee Se Hee, Lee Jae Ha, Lee Ho Won, Kim Yang-Hoon, Jeong Ok Chan, Ahn Ji-Young

机构信息

Department of Microbiology, Chungbuk National University, 1 Chungdae-Ro, Seowon-Gu, Cheongju 28644, South Korea.

Graduate School of Mechanical Engineering, Inje University, 197 Inje-ro, Gimhae, Gyungnam 621-749, South Korea.

出版信息

PLoS One. 2016 Jul 22;11(7):e0159777. doi: 10.1371/journal.pone.0159777. eCollection 2016.

Abstract

We fabricated droplet-based microfluidic platform for copolymerizable microspheres with acrydite modified DNA probe. The copolymerizable 3-D polyacrylamide microspheres were successfully produced from microcontinuous-flow synthesis with on-channel solidification. DNA copolymerization activity, surface presentation and thermostability were assessed by using fluorescent labeled complementary probe. The binding performance was only visible on the surface area of oligo-microspheres. We show that the resulting oligo-microspheres can be directly integrated into a streamlined microsphere-PCR protocol for amplifying ssDNA. Our microspheres could be utilized as a potential material for ssDNA analysis such as DNA microarray and automatic DNA SELEX process.

摘要

我们构建了基于液滴的微流控平台,用于制备带有丙烯酰胺修饰DNA探针的可共聚微球。通过微连续流合成和通道内固化成功制备了可共聚的三维聚丙烯酰胺微球。利用荧光标记的互补探针评估了DNA共聚活性、表面呈现和热稳定性。结合性能仅在寡聚微球的表面积上可见。我们表明,所得的寡聚微球可直接整合到用于扩增单链DNA的简化微球-PCR方案中。我们的微球可作为单链DNA分析的潜在材料,如DNA微阵列和自动DNA SELEX过程。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/031e/4957773/5f70b4ceea8e/pone.0159777.g001.jpg

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