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SRX7917437: GSM4413431: AD_60_Biomek_H3K27me3; Drosophila melanogaster; ChIP-Seq
1 ILLUMINA (Illumina NovaSeq 6000) run: 3.5M spots, 432.4M bases, 233.9Mb downloads

Submitted by: NCBI (GEO)
Study: AutoRELACS: Automated Generation And Analysis of Ultra-parallel ChIP-seq
show Abstracthide Abstract
We present AutoRELACS, an automated implementation of the RELACS protocol using the Biomek i7 automated workstation (Beckman&Coulter). We test the performance of AutoRELACS by assessing 1) the scalability of the chromatin barcode integration step, 2) the quality of the generated data in comparison to the benchmark set by the manual protocol, and 3) the sensitivity of the automated method when working with low (= 25.000 cells/sample) and very low (= 5.000 cells/sample) cell numbers. Overall design: 1) First, we test the effciency of automated barcode integration. We barcode 60 batches of S2 cells and we sequence only the input chromatin. 2) To test the overall quality of AutoRELACS in comparison to the manual protocol, we barcode the chromatin of 28 batches of S2 cells using custom RELACS adaptors and we pool them together using Biomek i7 automated workstation. The chromatin is used to generate genome-wide profiles for H3K4me3, H3K27ac and H3K27me3. 3) To test the sensitivity of the automated RELACS protocol, we profile the same three histone marks using 4 batches of HepG2 cells. We distribute the barcoded cells into two poolings. The Low pool contained a total of 300,000 cells, while the Very Low pool contained a total of 60,000 cells (see Fig 3a of publication for details).
Sample: AD_60_Biomek_H3K27me3
SAMN14383711 • SRS6323822 • All experiments • All runs
Library:
Instrument: Illumina NovaSeq 6000
Strategy: ChIP-Seq
Source: GENOMIC
Selection: ChIP
Layout: PAIRED
Construction protocol: HepG2 and S2 cells were fixed in 1% methanol-free formaldehyde (Thermo Scientific, 28906) in D-MEM (for HepG2 cells) or Express Five SFM (for S2 cells) for 15 min at room temperature under gentle shaking. Formaldehyde was quenched for 5 min by adding 125 mM glycine final concentration. Cells were rinsed twice with ice-cold PBS, harvested by scraping (HepG2) and pelleted (300 g, 10 min, 4 °C). Cells were processed according to RELACS / AutoRELACS protocols Libraries were prepared according to Illumina's instructions using NebNext Ultra II kit. After Illumina adapter ligation, library was PCR amplified. Fragments were size-selected to eliminate adapter dimers contamination using AMPure XP beads. The purified DNA was captured on an Illumina flow cell for cluster generation. Libraries were sequenced on NovaSeq 6000 machine, following manufacturer's protocol. High-throughput RELACS ChIP-seq and automated high-throughput AutoRELACS ChIP-seq
Experiment attributes:
GEO Accession: GSM4413431
Links:
Runs: 1 run, 3.5M spots, 432.4M bases, 233.9Mb
Run# of Spots# of BasesSizePublished
SRR113130183,487,267432.4M233.9Mb2020-07-28

ID:
10352372

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