Surface modification of poly(methyl methacrylate) microfluidic devices for high-resolution separations of single-stranded DNA.
While polymer-based microfluidic devices offer some unique opportunities in developing low-cost systems for a variety of application areas, the ability to sort electrophoretically with high efficiency a number of different targets has remained somewhat elusive with an example consisting of achieving single base resolution as required for DNA sequencing. While the reasons for this are many-fold, it is clear that some type of coating is required on the polymer substrate to suppress the EOF and/or minimize potential solute/wall interactions. To this end, we report on a simple grafting procedure to allow the formation of polymer coats, which in this example used linear polyarcylamides (LPAs), onto a poly(methyl methacrylate) (PMMA) microfluidic device. The procedure involved creating an amine-terminated PMMA surface by appropriately functionalizing the PMMA through either a chemical or photochemical process. The aminated surface could then be used to covalently anchor methacrylic acid, which was used as a scaffold to produce LPAs on the surface through radical polymerization of acrylamide. The resulting surfaces demonstrated EOFs that were nearly an order of magnitude smaller than native PMMA. In addition, these LPA-coated devices could produce highly reproducible migration times of over approximately 20 runs with plate numbers exceeding 10(5) m(-1). Using gel electrophoretic analysis of a single base track generated from an M13mp18 template using Sanger cycle sequencing and dye-primer chemistry, the resolution value obtained for bases 199 and 200 was 0.18 while for bases 208 and 209 it was 0.21. For the native PMMA, these bands were found to comigrate.
Llopis
Shawn L
SL
Department of Chemistry, Department of Mechanical Engineering, Center for BioModular Multi-Scale Systems, Louisiana State University, Baton Rouge, LA 70803, USA.
Osiri
John
J
Soper
Steven A
SA
D004277:DNA, Single-Stranded / Q000302:isolation & purification*; D053841:Electroosmosis; D020366:Methylmethacrylate / Q000737:chemistry*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D017422:Sequence Analysis, DNA / Q000295:instrumentation*; D013499:Surface Properties
D004277:DNA, Single-Stranded; D020366:Methylmethacrylate