Single molecule analysis of bacterial 16s rdna pcr products in submicrometer fluidic channels

AGRO 118

Samuel Stavis, School of Applied and Engineering Physics, Cornell University, Clark Hall, Ithaca, NY 14853, Stéphane C. Corgié, Department of Biological and Environmental Engineering, Cornell Unviersity, Riley-Robb Hall, Ithaca, NY 14853, Benjamin R. Cipriany, School of Applied and Engineering Physics, Cornell Unviersity, Clark Hall, Ithaca, NY 14853, H. G. Craighead, hgc1@cornell.edu, School of Engineering and Applied Physics, Cornell University, Ithaca, NY 14853, and Larry P. Walker, Department of Agricultural and Biological Engineering, Cornell University, Ithaca, NY 14850.
The manipulation, detection and analysis of nucleic acids have become essential tasks in many biological applications ranging from genomic sequencing to microbial ecology. The analytical demands of these applications have stimulated the development of Lab-on-a-Chip (LoaC) and Micro-Total-Analysis-System (ƒÝTAS) technology with integrated nanostructures to increase sensitivity and throughput. Laser induced fluorescence in submicrometer fluidic channels was used to analyze Polymerase Chain Reaction (PCR) products from the 16S rDNA of Thermobifida fusca (325 bp). Oligonucleotide universal bacterial primers and PCR amplicons were detected at all phases of the reaction without post-amplification purification or size screening. Primers were fluorescently labeled with single Alexa Fluor 488 or Alexa Fluor 594 fluorophores, resulting in double labeled amplicons.Photon burst analysis was used to detect and identify individual primers and amplicons, and fluorescence correlation and crosscorrelation spectroscopy were used to account for analyte flow speed.
 

Nanotechnology in Agriculture
8:30 AM-11:40 AM, Wednesday, August 22, 2007 BCEC -- 259A, Oral

Division of Agrochemicals

The 234th ACS National Meeting, Boston, MA, August 19-23, 2007