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Sample GSM1326467 Query DataSets for GSM1326467
Status Public on Feb 23, 2014
Title HAEC input TSA [MBD-seq]
Sample type SRA
Source name HAEC input TSA
Organism Homo sapiens
Characteristics cell type: Primary Human Aortic Endothelial Cells (HAECs)
passages: Passage 4-6
treated with: 500 nM TSA for 12hrs
Treatment protocol HAEC were incubated for 12 hours with 500 nM TSA.
Growth protocol Primary HAECs (human aortic endothelial cells), purchased from Lonza (Walkersville, MD, USA), were cultured in EBM-2 medium (Lonza) containing EGM-2 growth factors and supplements (Lonza) and 10% fetal bovine serum (Gibco).
Extracted molecule genomic DNA
Extraction protocol Genomic DNA derived from treated and controls HAECs was fragmented to 150-200bp by Diagnode sonicator and subjected to methylated DNA enrichment using MethylMiner Kit (Invitrogen, ME10035) in triplicate. Briefly, methylated DNA was captured from 500ng of fragmented genomic DNA via binding MBD2 protein coupled to magnetic Dynabeads, which was incubated for 1hr with rotation. The methylated fragments were then eluted in three distinct subpopulation using 0.6M, 1M and 2M salt elution buffers respectively (supplied in MethylMiner kit). Methylated DNA fragments, eluted with 2M salt buffer, were purified and selected for sequencing on the Illumina Genome Analyser.
Libraries were prepared according to the protocols described in the Illumina (San Diego, CA, US) ChIP-seq library preparation kit (cat# IP-102-1001).
Library strategy MBD-Seq
Library source genomic
Library selection MBD2 protein methyl-CpG binding domain
Instrument model Illumina Genome Analyzer IIx
Description HAEC_input_TSA_FC046_7
Data processing Illumina pipeline 1.4 was used for base calling
Reads were aligned to the hg19 genome using bwa 0.5.9 with default parameters
A set of peaks were generated for each sample vs Input using MACS 1.4.1
Genome_build: hg19
Supplementary_files_format_and_content: bed files comparing sample vs Input
Submission date Feb 12, 2014
Last update date May 15, 2019
Contact name Assam El-Osta
Organization name Baker Heart and Diabetes Institute
Lab Human Epigenetics
Street address 75 Commercial Road
City Melbourne
ZIP/Postal code 3004
Country Australia
Platform ID GPL10999
Series (2)
GSE37378 Vascular histone deacetylation by pharmacological HDAC inhibition
GSE54911 Vascular histone deacetylation by pharmacological HDAC inhibition [TSA, MBD-seq]
BioSample SAMN02640494
SRA SRX469928

Supplementary data files not provided
SRA Run SelectorHelp
Raw data are available in SRA
Processed data not provided for this record

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