Towards dynamic coating of glass microchip chambers for amplifying DNA via the polymerase chain reaction.
As microchip technology evolves to allow for the integration of more complex processes, particularly the polymerase chain reaction (PCR), it will become necessary to define simple approaches for minimizing the effects of surfaces on the chemistry/processes to be performed. We have explored alternatives to silanization of the glass surface with the use of additives that either dynamically coat or adsorb to the glass surface. Polyethylene glycol, polyvinylpyrrolidone (PVP), and hydroxyethylcellulose (HEC) have been explored as potential dynamic coatings and epoxy (poly)dimethylacrylamide (EPDMA) evaluated as an adsorbed coating. By carrying out analysis of the PCR products generated under different conditions via microchip electrophoresis, we demonstrate that these coating agents adequately passivate the glass surface in a manner that prevents interference with the subsequent PCR process. While several of the agents tested allowed for PCR amplification of DNA in glass, the EPDMA was clearly superior with respect to ease of preparation. However, more efficient PCR (larger mass of amplified product) could be obtained by silanizing the glass surface.
Giordano
B C
BC
Department of Chemistry, University of Virginia, Charlottesville 22901, USA.
Copeland
E R
ER
Landers
J P
JP
D000180:Acrylic Resins; D010582:Bacteriophage lambda / Q000737:chemistry; D002021:Buffers; D002482:Cellulose / Q000031:analogs & derivatives*; D020099:Coated Materials, Biocompatible; D004279:DNA, Viral / Q000032:analysis; D004852:Epoxy Compounds; D004867:Equipment Design; D005898:Glass; D007202:Indicators and Reagents; D008832:Microchemistry / Q000295:instrumentation*; D008904:Miniaturization; D016133:Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods; D011205:Povidone / Q000031:analogs & derivatives*; D012821:Silanes; D012997:Solvents; D013499:Surface Properties
D000180:Acrylic Resins; D002021:Buffers; D020099:Coated Materials, Biocompatible; D004279:DNA, Viral; D004852:Epoxy Compounds; D007202:Indicators and Reagents; D012821:Silanes; D012997:Solvents; C413461:poly(dimethylacrylamide-co-allyl glycidyl ether); C077842:polyvinylpolypyrrolidone; D002482:Cellulose; C002283:hydroxyethylcellulose; D011205:Povidone