Optimization of parameters for the automated microfluidic ChIP protocol. ChIP-qPCR are depicted on a positive (Actb) and a negative locus, with H3K4me3 recoveries plotted with ±SEM. (A) Recoveries using various types of frit layer composition that allow packing of antibody binding beads in the microfluidic reactors. (B) Recoveries using various column sizes (Supplemental Fig. S4A) built using different amounts of antibody binding beads. (C) Recoveries of various types of beads used to construct the antibody binding column. (D) Recoveries using varying chromatin loading pressures. (E) Final recoveries with optimized parameters compared with initial testing. (F) Recoveries of conventional versus low-input automated microfluidic ChIP-qPCR, showing high yields and reproducibility of microfluidic ChIP compared with conventional chip.
