Development of an entangled polymer solution for improved resolution in DNA analysis using a portable microfluidic instrument

CHED 298

Oscar G. Cabrices, ocabr001@fiu.edu, Kerry Opel, and Bruce R. McCord, mccordb@fiu.edu. Department of Chemistry, Florida International University, University Park 11200 SW 8th Street, Miami, FL 33199
The purpose of this research is to develop an entangled polymer solution for short tandem repeat (STR) separations on microfluidic devices. STRs are genomic elements consisting of 2-7 basepair sequences repeated various number of times. Recently a number of commercial systems have appeared using short fluidic channels for the rapid detection of native PCR amplified DNA using laser induced fluorescence. To be applicable for STR analysis these systems require improved separation efficiency and /or redesign due to their shorter separation channels. Currently, the resolution for the commercial DNA chip systems is approximately 5 bp. To improve resolution we are examining the application of polymeric buffers in combination with denaturants. The buffer utilized in this research includes Polyvinylpyrrolidone (PVP) (MW= 1,000,000 g/mol) in Tris-Borate-EDTA (TBE) buffer, with a urea denaturant for single stranded DNA analysis. Initial results in standard CE systems show good reproducibility for this polymer with minimal osmotic flow.