H3K27me3 Antibody, SNAP-Certified™ for CUT&RUN and CUT&Tag

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Figure 2: CUT&RUN genome-wide enrichment
CUT&RUN was performed as described above. Sequence reads were aligned to 18,793 annotated transcription start sites (TSSs ± 2 kbp). Signal enrichment was sorted from highest to lowest (top to bottom) relative to the H3K4me3 - 500k cells sample (all gene rows aligned). High, medium, and low intensity are shown in red, yellow, and blue, respectively. H3K4me3 positive control and H3K27me3 antibodies produced the expected diametric enrichment pattern, which was consistent between 500k and 50k cells and greater than the IgG negative control.

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Figure 3: H3K27me3 CUT&RUN representative browser tracks
CUT&RUN was performed as described above. Gene browser shots were generated using the Integrative Genomics Viewer (IGV, Broad Institute). H3K27me3 antibody tracks display the characteristic broad, intergenic enrichment known to be consistent with the function of this PTM. Similar results in peak structure and location were observed for both 500k and 50k cell inputs.

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CUT&RUN methods
CUT&RUN was performed on 500k and 50k K562 cells with the SNAP-CUTANA™ K-MetStat Panel (EpiCypher 19-1002) spiked-in prior to the addition of 0.5 µg of either IgG negative control (EpiCypher 13-0042), H3K4me3 positive control (EpiCypher 13-0041), or H3K27me3 antibodies. The experiment was performed using the CUTANA™ ChIC/CUT&RUN Kit v3.0 (EpiCypher 14-1048). Library preparation was performed with 5 ng of CUT&RUN enriched DNA (or the total amount recovered if less than 5 ng) using the CUTANA™ CUT&RUN Library Prep Kit (EpiCypher 14-1001/14-1002). Both kit protocols were adapted for high throughput Tecan liquid handling. Libraries were run on an Illumina NextSeq2000 with paired-end sequencing (2x50 bp). Sample sequencing depth was 16.8 million reads (IgG 500k cell input), 14.4 million reads (H3K4me3 500k cell input), 16.4 million reads (H3K27me3 500k cell input) and 11.6 million reads (H3K27me3 50k cell input). Data were aligned to the hg19 genome using Bowtie2. Data were filtered to remove duplicates, multi-aligned reads, and blacklist regions.