|
| Status |
Public on Nov 27, 2019 |
| Title |
H18neg_ATAC_R1 |
| Sample type |
SRA |
| |
|
| Source name |
H18neg_ATAC_R1
|
| Organism |
Gallus gallus |
| Characteristics |
strain: Wild-type
development: Embryo
development: HH18
tissue: Negative control (NC2-, E2- cells)
isolation_method: Dissociation and FACS
treatment: NC2:citrine, E2:citrine electroporation at HH8/9
amplification: NEB Next High-Fidelity 2X PCR Master Mix
assay: ATAC-seq
|
| Treatment protocol |
Dissected vagal regions from electroporated embryos were dissociated with dispase (1.5mg/ml in DMEM/10mM Hepes pH 7.5) at 37°C for 15min with intermittent pipetting to achieve a single cell suspension and 0.05% Trypsin at 37°C for a final 3min. The reaction was stopped and cells were resuspended in an excess of Hanks buffer. Cells were centrifuged at 500g for 10min and resuspended in Hanks buffer, passed through a 0.22mm filter and centrifuged at 750g for 10min, pelleted cells were resuspended in 500µl Hanks buffer. Fluorescent positive cells were sorted and collected using BD FACS-Aria Fusion.
|
| Growth protocol |
Fertilised wild-type chicken eggs were obtained from Henry Stewart & Co (Norfolk), staged according to Hamburger and Hamilton (1951) (24). All experiments were performed on chicken embryos younger than 12 days of development, and as such were not regulated by the Animals (Scientific Procedures) Act 1986.
|
| Extracted molecule |
genomic DNA |
| Extraction protocol |
FAC-sorted cells were lysed (10mM Tris-HCl, pH7.4, 10mM NaCl, 3mM MgCl2, 0.1% Igepal) and tagmented using Illumina Nextera DNA kit (FC-121-1030) for 20mins at 37°C. Tagmented DNA was amplified using NEB Next High-Fidelity 2X PCR Master Mix for 11 cycles. Tagmentation efficiency was assessed using Agilent Tapestation. Three biological replicates were obtained for each stage.
Libraries were prepared using Illumina Nextera DNA kit and sequenced on Illumina NextSeq500 using 40bp paired end reads
|
| |
|
| Library strategy |
ATAC-seq |
| Library source |
genomic |
| Library selection |
other |
| Instrument model |
Illumina NextSeq 500 |
| |
|
| Description |
Wild-type
ATAC_featurecounts_raw.txt
|
| Data processing |
Reads were mapped using Bowtie (v.1.0.0) to galGal5.
Peak calling was performed using MACS2, de novo motif search was performed using Homer (v.4.7)
Genome_build: galGal5
|
| |
|
| Submission date |
Jan 27, 2019 |
| Last update date |
Nov 27, 2019 |
| Contact name |
Tatjana Sauka-Spengler |
| E-mail(s) |
[email protected]
|
| Organization name |
MRC Weatherall Institute of Molecular Medicine
|
| Department |
University of Oxford
|
| Lab |
Sauka-Spengler lab
|
| Street address |
Radcliffe Department of Medicine
|
| City |
Oxford |
| State/province |
Oxfordshire |
| ZIP/Postal code |
OX3 9DS |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL19787 |
| Series (2) |
| GSE125707 |
Early chromatin shaping predetermines multipotent vagal neural crest into neural, neuronal and mesenchymal lineages |
| GSE125711 |
Early chromatin shaping predetermines multipotent vagal neural crest into neural, neuronal and mesenchymal lineages |
|
| Relations |
| BioSample |
SAMN10826860 |
| SRA |
SRX5296622 |
