Automated Microfluidic Platform for Serial Polymerase Chain Reaction and High-Resolution Melting Analysis

Weidong Cao, Brian Bean, Scott Corey, Johnathan S. Coursey, Kenton C. Hasson, Hiroshi Inoue, Taisuke Isano, Sami Kanderian, Ben Lane, Hongye Liang, Brian Murphy, Greg Owen, Nobuhiko Shinoda, Shulin Zeng, Ivor T. Knight

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

We report the development of an automated genetic analyzer for human sample testing based on microfluidic rapid polymerase chain reaction (PCR) with high-resolution melting analysis (HRMA). The integrated DNA microfluidic cartridge was used on a platform designed with a robotic pipettor system that works by sequentially picking up different test solutions from a 384-well plate, mixing them in the tips, and delivering mixed fluids to the DNA cartridge. A novel image feedback flow control system based on a Canon 5D Mark II digital camera was developed for controlling fluid movement through a complex microfluidic branching network without the use of valves. The same camera was used for measuring the high-resolution melt curve of DNA amplicons that were generated in the microfluidic chip. Owing to fast heating and cooling as well as sensitive temperature measurement in the microfluidic channels, the time frame for PCR and HRMA was dramatically reduced from hours to minutes. Preliminary testing results demonstrated that rapid serial PCR and HRMA are possible while still achieving high data quality that is suitable for human sample testing.

Original languageEnglish (US)
Pages (from-to)402-411
Number of pages10
JournalJournal of Laboratory Automation
Volume21
Issue number3
DOIs
StatePublished - Jun 2016

All Science Journal Classification (ASJC) codes

  • Computer Science Applications
  • Medical Laboratory Technology

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