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Sample GSM6475138 Query DataSets for GSM6475138
Status Public on May 24, 2023
Title CT26 Phf8 KO2_H3K9me2
Sample type SRA
 
Source name CT26
Organism Mus musculus
Characteristics cell line: CT26
cell type: colorectal cancer
genotype: CT26 Phf8 knockout cell
chip antibody: anti-H3K9me2 (ab176882, abcam)
Extracted molecule genomic DNA
Extraction protocol Cells were fixed with 1% formaldehyde for 10 minutes. After quenching and cell lysis, chromatin was sonicated using a E220 focused-ultrasonicator (Covaris).
ChIP-seq libraries were prepared using the KAPA HTP Library Preparation Kit complemented with NEXTflex DNA Barcodes from Bioo Scientific. 10 ng of DNA was used as starting material for input and ip samples. Libraries were amplified using 13 cycles on the thermocycler. Post amplification libraries were size selected at 250-450bp in length using Agencourt AMPure XP beads from Beckman Coulter. Libraries were validated using the Agilent High Sensitivity DNA Kit.
 
Library strategy ChIP-Seq
Library source genomic
Library selection ChIP
Instrument model Illumina NovaSeq 6000
 
Data processing Basecalls were performed using bcl2fastq v2.17 for Novaseq output.
ChIP-seq reads were trimmed from 3' end until the final base had a quality score > 30, using Trimmomatic v0.33, discarding reads left with < 20 bp
ChIP-seq reads were aligned to the UCSC mm10 genome using Bowtie version 1.1.2. Only uniquely mapping reads with at most two mismatches were retained.
To create ChIP-seq coverage plots, the locations of the mapped ChIP-seq reads were extended to 150 bp to represent sequenced fragments, renormalized (to reads per million, rpm) and reformatted in the bigWig file format.
Peaks were called uing MACS v2.1.0 with the significance cut-off q-value <=0.01
bigWig files were generated using the genomicRanges R package. Score represents the normalized coverage of DNA fragments at a given genomic coordinate. narrowPeak files were generated using MACS v2 with default settings.
Assembly: mm10
Supplementary files format and content: bigWig, narrowPeak (except for Input sample)
 
Submission date Aug 18, 2022
Last update date May 25, 2023
Contact name Yanan Liu
E-mail(s) Lyn19950902@outlook.com
Organization name East China Normal University
Department School of Life Sciences
Street address 500 Dongchuan Road, Minhang District, Shanghai, China
City China
ZIP/Postal code 200241
Country China
 
Platform ID GPL24247
Series (2)
GSE211524 PHF8 enables immune evasion by silencing endogenous retroelements [CHIP-seq]
GSE211526 PHF8 enables immune evasion by silencing endogenous retroelements.
Relations
BioSample SAMN30383409
SRA SRX17132127

Supplementary file Size Download File type/resource
GSM6475138_CT26_Phf8_KO2_H3K9me2.bw 171.6 Mb (ftp)(http) BW
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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