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Sample GSM5099941 Query DataSets for GSM5099941
Status Public on Aug 30, 2021
Title WRKY12
Sample type SRA
 
Source name seedling
Organism Triticum urartu
Characteristics tissue: seedling
dap antibody: Halo (Promega-G7281)
accession: G1812 (PI428198)
Growth protocol 16h light, 8h dark, 22°C
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).
 
Library strategy OTHER
Library source genomic
Library selection other
Instrument model Illumina NovaSeq 6000
 
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
 
Submission date Feb 22, 2021
Last update date Aug 30, 2021
Contact name yijing zhang
E-mail(s) zhangyijing@fudan.edu.cn
Organization name Fudan University
Department Biochemistry
Lab Functional Epigenomics Group
Street address 2005 Songhu Road
City shanghai
ZIP/Postal code 200438
Country China
 
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
Relations
BioSample SAMN18026244
SRA SRX10149605

Supplementary file Size Download File type/resource
GSM5099941_WRKY12.bw 26.9 Mb (ftp)(http) BW
GSM5099941_WRKY12_peaks.txt.gz 1.3 Mb (ftp)(http) TXT
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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