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Sample GSM4800067 Query DataSets for GSM4800067
Status Public on Sep 24, 2020
Title H3K4me1 from UC Davis FAANG pig liver rep P350
Sample type SRA
 
Source name Liver
Organism Sus scrofa
Characteristics chip antibody: H3K4me1 (Diagenode,#C15410037,lot A1862D)
tissue: liver
Extracted molecule genomic DNA
Extraction protocol ChIP-seq experiments were performed on frozen tissue using the iDeal ChIP-seq kit for Histones (Diagenode Cat.#C01010059, Denville, NJ) according to the manufacturer’s protocol except for the following changes. 20-30 mg of frozen tissue was powdered using liquid nitrogen in pre-chilled mortar. Cross-linking was performed with 1% formaldehyde which was diluted from 16% methanol-free formaldehyde (Thermo Scientific, Cat.#28906, Waltham, MA) for 8 minutes and quenched with glycine for 10 minutes. Nuclei were harvested by centrifugation at 2000g for 5 minutes and resuspended in iS1 buffer for incubation on ice for 30 minutes. Chromatin was sheared using the Covaris E220 between 6-12 minutes depending on the tissue. For immunoprecipitation experiments, about 1000 ng of sheared chromatin (estimated from DNA extraction) was used as input after which the kit protocol was followed with 1 μg (histone modifications) or 1.5 μg (CTCF) of antibody. The following antibodies used were from Diagenode: H3K4me3 (in kit), H3K27me3 (#C15410069), H3K27ac (#C15410174), H3K4me1 (#C15410037), and CTCF (#15410210). An input (no antibody) was performed for each sample.
NEBNext Ultra DNA library prep kit for Illumina libraries (New England Biolabs #E7645L, Ipswich, MA) was used for library construction, selecting for 150-200 bp (H3K4me3, H3K27ac, CTCF) or 200-400 bp (H3K27me3, H3K4me1) insert fragment sizes using Ampure beads (Beckman Coulter #A63881). Libraries were sequenced on Illumina’s HiSeq 4000 with single-end 50 bp reads.
 
Library strategy ChIP-Seq
Library source genomic
Library selection ChIP
Instrument model Illumina HiSeq 4000
 
Description SAMEA4454578
Data processing Reads were trimmed with Trim Galore! 0.4.5 with the default arguments
Reads were aligned with BWA mem 0.7.17 to the galGal6, Sscrofa11.2, and ARS-UCD1.2 genomes
Alignments were filtered with Samtools 1.10 using arguments -F 1804 -q 30
Duplicates were removed with Picard toolkit 2.18.17
Genome_build: galGal6,susScr11,bosTau9
Supplementary_files_format_and_content: bam file, aligned reads generated by data processing steps
Supplementary_files_format_and_content: bed file, peak calls using Macs 2.1.1 -q 0.01
 
Submission date Sep 23, 2020
Last update date Sep 24, 2020
Contact name Colin Kern
Organization name UC San Diego
Street address 9500 Gilman Dr
City La Jolla
State/province CA
ZIP/Postal code 92093
Country USA
 
Platform ID GPL22475
Series (2)
GSE158427 Comparative functional genome annotation of livestock species identifies core amniote regulatory features and avian enhancer versatility (H3K4me1 ChIP-Seq)
GSE158430 Comparative functional genome annotation of livestock species identifies core amniote regulatory features and avian enhancer versatility
Relations
BioSample SAMN16245661
SRA SRX9177184

Supplementary file Size Download File type/resource
GSM4800067_H3K4me1_Liver_P350_Peaks.bed.gz 2.7 Mb (ftp)(http) BED
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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