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SRX4116033: GSM3149070: B2_ER_FA; Homo sapiens; ChIP-Seq
1 ILLUMINA (Illumina HiSeq 2500) run: 24.1M spots, 1.6G bases, 435.1Mb downloads

Submitted by: NCBI (GEO)
Study: Optimized ChIP-seq procedure facilitates hormone receptor profiling in human tumors
show Abstracthide Abstract
Performing ChIP-seq analyses in clinical specimens has remained largely challenging due to multiple technical limitations and low quantities of starting material, resulting in low enrichments and poor signal-to-noise ratio. Here, we refined the original protocols for transcription factor ChIP-seq analyses in breast, prostate, and endometrial tumor tissue. In addition to the standard fixative formaldehyde, a second crosslinker Disuccinimidyl glutarate (DSG) was included in the procedure. Overall design: AR, FOXA1, ERa, H3K27ac & H3K4me3 ChIP-seq data with and without double croslinking in celllines (LNCAP & MCF-7) and in human tissues (Prostate, Breast, Endometrium and Prostate samples from core needle biopsies).
Sample: B2_ER_FA
SAMN09238370 • SRS3331324 • All experiments • All runs
Organism: Homo sapiens
Library:
Instrument: Illumina HiSeq 2500
Strategy: ChIP-Seq
Source: GENOMIC
Selection: ChIP
Layout: SINGLE
Construction protocol: Chromatin immunoprecipitations were performed as described previously with minor changes (Zwart et al., 2013). Samples were crosslinked in solution A with 2mM DSG (CovaChem) for 25 minutes at room temperature. After 25 minutes, 1% formaldehyde was added for 20 minutes and subsequently quenched with glycine. Samples were lysed as described (Schmidt et al., 2009) and sonicated for at least 10 cycles of 30s on, 30s off using a Diagenode Bioruptor Pico. For each ChIP, 5ug of antibody was conjugated with 50ul Protein A magnetic beads. Antibodies used were AR (sc-816, Santa Cruz), H3K27ac (39133, Active Motif), H3K4me3 (Ab8580, Abcam) and H3K27me3 (39155, Active Motif). Immunoprecipitated DNA was processed for library preparation (Part# 0801-0303, KAPA biosystems kit). Immunoprecipitated DNA was processed for library preparation (Part# 0801-0303, KAPA biosystems kit).
Experiment attributes:
GEO Accession: GSM3149070
Links:
Runs: 1 run, 24.1M spots, 1.6G bases, 435.1Mb
Run# of Spots# of BasesSizePublished
SRR720704324,101,1801.6G435.1Mb2019-01-01

ID:
5601594

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