The rotary zone thermal cycler: a low-power system enabling automated rapid PCR

PLoS One. 2015 Mar 31;10(3):e0118182. doi: 10.1371/journal.pone.0118182. eCollection 2015.

Abstract

Advances in molecular biology, microfluidics, and laboratory automation continue to expand the accessibility and applicability of these methods beyond the confines of conventional, centralized laboratory facilities and into point of use roles in clinical, military, forensic, and field-deployed applications. As a result, there is a growing need to adapt the unit operations of molecular biology (e.g., aliquoting, centrifuging, mixing, and thermal cycling) to compact, portable, low-power, and automation-ready formats. Here we present one such adaptation, the rotary zone thermal cycler (RZTC), a novel wheel-based device capable of cycling up to four different fixed-temperature blocks into contact with a stationary 4-microliter capillary-bound sample to realize 1-3 second transitions with steady state heater power of less than 10 W. We demonstrate the utility of the RZTC for DNA amplification as part of a highly integrated rotary zone PCR (rzPCR) system that uses low-volume valves and syringe-based fluid handling to automate sample loading and unloading, thermal cycling, and between-run cleaning functionalities in a compact, modular form factor. In addition to characterizing the performance of the RZTC and the efficacy of different online cleaning protocols, we present preliminary results for rapid single-plex PCR, multiplex short tandem repeat (STR) amplification, and second strand cDNA synthesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Automation, Laboratory*
  • Humans
  • Polymerase Chain Reaction / methods*
  • Polymerase Chain Reaction / standards
  • Reproducibility of Results
  • Sensitivity and Specificity

Grants and funding

This work was funded in part by the U.S. Department of Defense Rapid Reaction Technology Office on behalf of the U.S. Army Criminal Investigation Laboratory as part of the Battlefield Automated DNA Analysis and Sampling System project. Additional support was provided by Sandia National Laboratories internal Laboratory Directed Research and Development grants. Support for Harrison Edwards, Daniel Lee, Caroline Moseley, and Karen Tew was provided by the Department of Homeland Security HS-STEM internship program. Sandia is a multiprogram laboratory operated by Sandia Corporation, a Lockheed Martin Company, for the United States Department of Energy under Contract DE-AC04-94AL85000. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.