GSM9439203: ASPC1 shZNF274 IgG rep1; Homo sapiens; OTHER
Cells were washed, bound to concanavalin A-coated magnetic beads (Bangs Laboratories, ca. no. BP531), permeabilized with Triton X-100, and bound with protein specific antibodies prior to submission to the Fred Hutchinson Cancer Center Genomics Shared Resource High Throughput Screening Lab for AutoCUT&RUN processing. Libraries were prepared using a Beckman Biomek i7 liquid handling instrument equipped with a 96S Super Magnet Plate (Alpaqua SKU A001322) for magnetic separation of samples during wash steps. End-repair, adapter ligation, and PCR amplification reactions were performed on a separate thermocycler, and purified PCR products were analyzed on an Agilent 4200 TapeStation. Gated size and quantification values were used to calculate sample library yields for pooling at equimolar concentration for paired-end 50x50 bp sequencing on an Illumina NovaSeq X Plus over one lane of a 10B-100 flow cell at the Fred Hutchinson Cancer Center Genomics Shared Resource.