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Status |
Public on Sep 04, 2020 |
Title |
YueLab-H3K27ac-embryonic-Brain-rep1 |
Sample type |
SRA |
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Source name |
Tissue
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Organism |
Danio rerio |
Characteristics |
strain: Tuebingen tissue: embryonic-Brain chip-seq antibody: H3K27ac antibody (Active Motif, 39155)
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Growth protocol |
Embryonic Tuebingen zebrafish were raised under standard laboratory conditions
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Extracted molecule |
genomic DNA |
Extraction protocol |
Zebrafish embryonic tissue ChIP-seq was performed similarly to the adult tissue with a few modifications. For embryonic trunk, 50 trunks of 1 dpf embryos were dissected and digested in 500 µL of 0.25% trypsin at room temperature for 20 min. The reaction was then neutralized with FBS. Trunk cells were washed once with cold 1X PBS and cross-linked with 1% formaldehyde at room temperature for 12 min. For embryonic neuronal cells, Tg(Huc:Kaede) transgenic fish were crossed with wildtype Tuebingen fish. At 1 dpf, embryos were checked under the fluorescence microscope and green positive embryos were selected, dechorionated and pipetted in calcium free Ringer's solution to remove yolk. Supernatant was removed and 500 µL of 0.25% trypsin was added to digeste embryos at room temperature for 20 min. Neutralized with 500 µL of FBS and washed once with cold 1X PBS, digested cells were then cross-linked with 1% formaldehyde at room temperature for 12 min. Cells were then sorted and only green fluoresced cells, which corresponded to embryonic neuronal cells, were collected . To prepare the library, eluted chromatin was reverse crosslinked and purified by phenol-chloroform extraction. Then DNA was end-repaired by END-IT DNA end-repair kit (Epicentre, ER81050) according to kit’s protocol, added A using Klenow fragment (3’->5’ exo-) (NEB, M0212S), ligated with Illumina TruSeq adaptor (Illumina, FC-121-3001) and subsequently amplified by PCR (Roche, kk2601). The quality and quantity of all the libraries were checked using BioAnalyzer High Sensitivity DNA Kit (Agilent). The libraries were sequenced on Illumina HiSeq 2500 with reads length of 2X 50 bp or 2X 100 bp.
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Library strategy |
ChIP-Seq |
Library source |
genomic |
Library selection |
ChIP |
Instrument model |
Illumina Genome Analyzer |
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Description |
YueLab-ChIP-seq-embryonic-Brain_H3K27ac_merged.narrowPeak
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Data processing |
RNA-seq reads were aligned to zv10 genome assembly using STAR; ChIP-seq and ATAC-seq reads were aligned to zv10 genome assembly using BWA HiC reads were aligned to zv10 genome assembly using Bowtie2 The TPM value of gene expression was caculated using RSEM ChIP-seq and ATAC-seq peaks were called using MACS2 with the following setting: ChIP-seq q-value <10e-2, p-value<10e-5, Change>1,FC>2. ATAC-seq: q-value<10e-2 and p-value<10e-5 HiC matrix was generated using HiC-Pro Genome_build: zv10 Supplementary_files_format_and_content: tab-delimited text files include TPM values for each Sample; the narrowPeak files included the peaks for each Sample; The .hic file were the matrix of Hi-C for each Sample.**All replicates were merged
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Submission date |
Jul 09, 2019 |
Last update date |
Sep 04, 2020 |
Contact name |
Yu Luan |
E-mail(s) |
yu.luan@northwestern.edu
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Organization name |
Northwestern University Feinberg School of Medicine
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Department |
Department of Biochemistry and Molecular Genetics
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Lab |
Feng Yue
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Street address |
303 E. Superior Simpson Querrey 7-518
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City |
CHICAGO |
State/province |
Illinois |
ZIP/Postal code |
60611 |
Country |
USA |
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Platform ID |
GPL10164 |
Series (1) |
GSE134055 |
A map of cis-regulatory elements and 3D genome structures in zebrafish |
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Relations |
BioSample |
SAMN12239167 |
SRA |
SRX6422934 |
Supplementary data files not provided |
SRA Run Selector |
Raw data are available in SRA |
Processed data are available on Series record |
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