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Status |
Public on Aug 30, 2021 |
Title |
Halo_rep3 |
Sample type |
SRA |
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Source name |
seedling
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Organism |
Triticum urartu |
Characteristics |
tissue: seedling dap antibody: Halo (Promega-G7281) accession: G1812 (PI428198)
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Growth protocol |
16h light, 8h dark, 22°C
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Extracted molecule |
genomic DNA |
Extraction protocol |
Genomic DNA was extracted from leaves using Plant DNAzol Reagent(invitrogen). RNA was extracted using TRIzol. Genomic DNA was fragmented. And then end repaired using the End-It kit(Lucigen) and A-tailed using Klenow(3'-5' exo-; NEB). Truncated Illumina Y-adapter was ligated to DNA using T4 DNA Ligase(Promega). Full length TF was cloned into pIX-Halo vector. Halo-tagged TF was expressed in vitro using TNT SP6 Coupled Wheat Germ Extract System(Promega). Halo-TF was immobilized by Magne HaloTag Beads(Promega) and then incubated with the DNA library. TF specific binding DNA was eluted for 10 min at 98°C and amplified with indexed Illumina primer using Phanta Max Super-Fidelity DNA Polymerase(Vazyme). Meanwhile, to capture background DNA which captured by Halo, pIX-Halo vector without TF cloned was expressed and incubated with the DNA library as well. The PCR product was purified using VAHTS DNA Clean Beads(Vazyme).
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Library strategy |
OTHER |
Library source |
genomic |
Library selection |
other |
Instrument model |
Illumina NovaSeq 6000 |
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Description |
Halo_merge.bw
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Data processing |
Library strategy: DAP-seq Sequencing reads were cleaned with the fastp(version 0.20.0) and Trim Galore (version 0.4.4), which eliminated bases with low quality scores(< 25) and irregular GC contents, sequencing adapters, and short reads. The remaining clean reads were mapped to the T. urartu genome with the Burrows–Wheeler Aligner(version 0.7.17-r1188). The MACS(version 2.2.6) program was used to identify the read-enriched regions(peaks) based on the criteria: P < 1e−10. The peaks detected from samples introduced with Halo tag only were considered as non-specific bindings, and TF peaks overlapping with peaks detected from Halo samples were removed for subsequent analysis. Genome_build: WheatTu IGDBv1.0 Supplementary_files_format_and_content: peak files and bigwig files
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Submission date |
Feb 22, 2021 |
Last update date |
Aug 30, 2021 |
Contact name |
yijing zhang |
E-mail(s) |
zhangyijing@fudan.edu.cn
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Organization name |
Fudan University
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Department |
Biochemistry
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Lab |
Functional Epigenomics Group
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Street address |
2005 Songhu Road
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City |
shanghai |
ZIP/Postal code |
200438 |
Country |
China |
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Platform ID |
GPL29754 |
Series (2) |
GSE167227 |
Evolutionary rewiring of wheat abiotic stress responsive network by lineage-specific transposable elements [DAP-Seq] |
GSE167229 |
Evolutionary rewiring of wheat abiotic stress responsive network by lineage-specific transposable elements |
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Relations |
BioSample |
SAMN18026272 |
SRA |
SRX10149620 |
Supplementary file |
Size |
Download |
File type/resource |
GSM5099956_Halo_rep3.bw |
29.8 Mb |
(ftp)(http) |
BW |
SRA Run Selector |
Raw data are available in SRA |
Processed data provided as supplementary file |
Processed data are available on Series record |
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