Effect of sieving polymer concentration on separation of 100 bp DNA Ladder by capillary gel electrophoresis

Abstract

We studied the effect of sieving polymer concentration on separation of a 100 bp DNA Ladder by capillary gel electrophoresis (CGE) using hydroxyethyl cellulose (HEC) with a molecular size of 1000 k. For measurement purposes, we selected a fused silica capillary with total length of 15 cm and effective length of 7.5 cm; this was applied to compact CGE equipment for a Point-Care-Testing (POCT) system. Measurement results of the 100 bp DNA Ladder sample indicated that small DNA separation was significantly affected by HEC sieving polymer concentration. This was due to the level of entanglement between small DNA molecules and the sieving polymer chain significantly influencing migration time, mobility, and resolution length of the CGE process. We concluded that 1.0 w/v % HEC sieving polymer concentration was optimal for CGE separation of DNA ≥1000bp in the 100 bp DNA Ladder (100-1500 bp) when using the short-length capillary.