DNA Separation by Microchip Electrophoresis Using Copolymers of Poly(vinylpyrrolidone) and Hydroxyethylcellulose

Abstract

Copolymers of poly(vinylpyrrolidone) (PVP) and hydroxyethylcellulose (HEC) were synthesized, with PVP to HEC molar ratios of 3∶1, 2∶1 and 1∶1. The copolymers were tested as separation media in DNA fragment separation analysis by microchip electrophoresis (MCE). Separation efficiency over 3.8×10 5 for 118 bp has been reached by using the bare channels without the additional polymer coating step. Under optimized separation conditions for longer read length DNA sequencing, the separation ability of the copolymers decreased with decreasing (PVP-co-HEC) molar ratio from 3∶1 to 2∶1 and 1∶1. In comparison with (PVP-co-HEC) 1∶1, the copolymer with (PVP-co-HEC) 3∶1 ratio showed high separation efficiency. By using a 20 g•L -1 copolymer with (PVP-co-HEC) 3∶1 ratio, ΦΧ174-HaeШ digest DNA marker was successfully separated within 3 min.

Multiplexed p53 Mutation Detection by Microchip Electro‐phoresis with Laser‐Induced Fluorescence Detector

“…19 A diode laser was used to generate an excitation beam at 488 nm and an incident power of about 4 mW. The laser beam was focused by a concave mirror, illuminating at 45° from below the chip, on the detection point of the chip channel, with a focus point of about 70 µm.…”