Detection of Low-Abundance KRAS Mutations in Colorectal Cancer Using Microfluidic Capillary Electrophoresis-Based Restriction Fragment Length Polymorphism Method with Optimized Assay Conditions
Figure 2
Optimization of separation parameters for the microfluidic capillary electrophoresis-based restriction fragment length polymorphism (µCE-based RFLP) platform.
(A) The effect of field strength. Conditions for separation were 2% HPC, 3×TBE with varied field strength. (B) The effect of polymer concentration. Conditions for separation were 3×TBE, 180 V/cm with varied polymer concentrations. (C) The effect of buffer concentration. Conditions for separation were 2% HPC, 180 V/cm with varied ionic strength. (D) µCE-based RFLP analysis of the products of enzymatic digestion. Products were separated in sieving buffers with 3×TBE, 2×TBE, or 1×TBE. Separations were performed using 2% HPC under a 180 V/cm electric field.