Picoliter DNA sequencing chemistry on an electrowetting-based digital microfluidic platform.

Published

Journal Article

The results of investigations into performing DNA sequencing chemistry on a picoliter-scale electrowetting digital microfluidic platform are reported. Pyrosequencing utilizes pyrophosphate produced during nucleotide base addition to initiate a process ending with detection through a chemiluminescence reaction using firefly luciferase. The intensity of light produced during the reaction can be quantified to determine the number of bases added to the DNA strand. The logic-based control and discrete fluid droplets of a digital microfluidic device lend themselves well to the pyrosequencing process. Bead-bound DNA is magnetically held in a single location, and wash or reagent droplets added or split from it to circumvent product dilution. Here we discuss the dispensing, control, and magnetic manipulation of the paramagnetic beads used to hold target DNA. We also demonstrate and characterize the picoliter-scale reaction of luciferase with adenosine triphosphate to represent the detection steps of pyrosequencing and all necessary alterations for working on this scale.

Full Text

Duke Authors

Cited Authors

  • Welch, ERF; Lin, Y-Y; Madison, A; Fair, RB

Published Date

  • February 2011

Published In

Volume / Issue

  • 6 / 2

Start / End Page

  • 165 - 176

PubMed ID

  • 21298802

Pubmed Central ID

  • 21298802

Electronic International Standard Serial Number (EISSN)

  • 1860-7314

International Standard Serial Number (ISSN)

  • 1860-6768

Digital Object Identifier (DOI)

  • 10.1002/biot.201000324

Language

  • eng