Entangled polymer networks continue to be of great interest as a means to separate single-stranded DNA fragments, particularly for DNA sequencing applications. While capillary electrophoresis instruments were very successful in sequencing the complete human genome as part of the Human Genome Project, the capital costs, required reagent volumes, and long run times make this technology infeasible for continued large-scale de novo sequencing projects. Microfluidic chips have shown tremendous promise for producing high quality DNA sequencing data while also lowering required reagent volumes and separation times. The ability to precisely control the injection plug width in the separation channel by specifying the microchip geometry enables shorter separation channels to be used, which result in shorter separation times. Performing DNA sequencing fragment separations in microchips, our group has previously described a poly(N,N-dimethylacrylamide) (pDMA) matrix and physically adsorbed poly(N-hydroxyethylacrylamide) wall coating system that enables ultra-fast DNA sequencing of 600 bases in approximately 6.5 minutes at 98.5% accuracy. This result is a 66% decrease in separation time over previously published results. To further explore this observed improvement in separation speed, we have varied pDMA matrix properties, such as polymer molar mass and polymer solution concentration, as well as several chip electrophoresis parameters, particularly separation length, temperature, and electric field strength, to determine the resulting sequencing read lengths and separation times. In this report, we discuss the effect each of these parameters has on both sequencing read length and separation time. We also discuss the effect that blending two pDMA samples of differing molar mass has on the resulting sequencing read lengths and separation time in microfluidic channels. These results will also be discussed in the context of different sequencing applications, specifically medical sequencing and full genome sequencing.