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Effect of tris(2-carboxyethyl)phosphine and tertiary butyl alcohol on the performance of convection polymerase chain reaction

  • Tae-Hoon Kim
  • Kyungyoung Song
  • Hyun Jin Hwang
  • Jeong Hee Kim
Original Article
  • 7 Downloads

Abstract

Rapid and on-site DNA-based molecular detection has become increasingly important for sensitive, specific, and timely detection and treatment of various diseases. To prepare and store biomolecule-containing reagents stably, reducing agents are used during protein preparation, and freeze-drying technology has been applied to the protein reagents. Some of the additives used during these processes may affect subsequent processes such as polymerase chain reaction (PCR). In this study, we evaluated the impact of TCEP, a reducing agent, and TBA, a freeze-drying medium, on the performance of convection PCR (cPCR) using a battery-operable PCR device. Singleplex cPCR detection of a 249 bp amplicon from human genomic DNA suggested that approximately 82% of performance was achieved in the presence of 0.1 mM TCEP and 1% TBA. The limit of detection and the minimum number of cycles at which amplicons began to appear was a little lower (~ 82% efficiency) or higher (20 vs 15 cycles), respectively, in the chemical-treated group than in the control group. With larger amplicons of 500 bp, the chemical-treated group revealed approximately 78% of performance and amplicons started to appear at 20 cycles of cPCR in both groups. Similar results were obtained with multiplex cPCR amplification.

Keywords

TCEP TBA cPCR On-site Molecular diagnostics 

Notes

Acknowledgements

This work was supported by Grant 10080151 from Korea Evaluation Institute of Industrial Technology, funded by the Ministry of Trade, Industry and Energy, Korea.

Compliance with ethical standards

Conflict of interest

The authors have declared that no competing interests exist.

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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Department of Life and Nanopharmaceutical Sciences, Graduate SchoolKyung Hee UniversitySeoulSouth Korea
  2. 2.Department of Oral Biochemistry and Molecular BiologyKyung Hee UniversitySeoulSouth Korea
  3. 3.R&D CenterAhram Biosystems IncSeoulSouth Korea
  4. 4.Department of Biochemistry and Molecular Biology, School of DentistryKyung Hee UniversitySeoulSouth Korea

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