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

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.

MeSH terms

  • DNA Probes
  • DNA Replication
  • DNA, Single-Stranded*
  • Microfluidics* / instrumentation
  • Microfluidics* / methods
  • Microspheres*
  • Oligonucleotide Probes*
  • Polymerization

Substances

  • DNA Probes
  • DNA, Single-Stranded
  • Oligonucleotide Probes

Grants and funding

The authors are grateful for the support of the "Cooperative Research Program for Agriculture Science & Technology Development (http://atis.rda.go.kr/rdais/main/main.vw, Project title: Development of Monitoring and Diagnostic method for Environmental Animal Disease, Project No. PJ010530)" Rural Development Administration, Republic of Korea). This research was also supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF, http://www.nrf.re.kr/nrf_eng_cms/) funded by the Ministry of Education (NRF-2013R1A1A2012537).