Consecutive Electrophoretic Separation of PCR Products Under a High-Ionic-Strength Solution on PMMA Chips with Enhanced Static Adsorptive Coat

Abstract

Consecutive polymerase chain reaction (PCR) product electrophoretic separation was done using a high-ionic-strength solution on poly(methyl methacrylate) (PMMA) micofluidic devices. Microchannels were modified with an enhanced static adsorptive coating method using 3% hydroxyethyl cellulose. The relative standard deviations of migration time and fluorescence intensity for the amplified cytosine deaminase PCR product of 258 bp (without pretreatments in 16 consecutive and rapid runs) on the PMMA chip were 0.88% and 3.5%, with a separation efficiency of 6.0 × 10⁵/m. PCR products were repeatedly separated on the modified chip without rinsing the channels with water and refilling the channels with the sieving matrix between runs.