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Sample GSM5100037 Query DataSets for GSM5100037
Status Public on Aug 30, 2021
Title G1812_RNAseq_G1812_CK_rep5
Sample type SRA
 
Source name seedling
Organism Triticum urartu
Characteristics tissue: seedling
accession: G1812 (PI428198)
Treatment protocol Regarding the cold and heat stress treatments, 7-day-old seedlings grown in soil were transferred to 4 °C for 5 hours, 40 °C for 7 hours, respectively. To assess the effects of drought, 7-day-old seedlings were cultivated in soil for another 2 weeks without watering. For the NaCl and ABA treatments, 7-day-old seedlings grown in soil were treated with 250 mM NaCl for 7 hours, 100 μm ABA for 2 weeks. For wounding stress, 7-day-old seedlings were injured on leaves by scissors and samples were taken 2.5 hours later.
Growth protocol 16h light, 8h dark, 22°C
Extracted molecule total RNA
Extraction protocol 2.2 μg DNA extracted from the harvested seedlings were used to prepare ChIP-seq, DNase-seq and Bisulphite-seq. RNA was extracted using TRIzol and 2μg total RNA were used to prepare RNA-seq. Nuclei were extracted with H1B buffer(20 mM Tris-HCl, pH 8.0, 50 mM EDTA, 5 mM spermidine, 0.15 mM spermine, 40% glycerol, and 0.1% mercaptoethanol) for DNase-seq.
Libraries were prepared for sequencing using standard Illumina protocols.
 
Library strategy RNA-Seq
Library source transcriptomic
Library selection cDNA
Instrument model Illumina NovaSeq 6000
 
Data processing Sequencing reads were cleaned with the Trim Galore, Trimmomatic, and Sickle programs.
The cleaned reads were mapped to WheatTu IGDBv1.0 using BWA for ChIP-seq and DNase-seq. HISAT2 for RNA-seq data. Bismark program for Bisulphite-seq.
For ChIP-seq and DNase-seq, MACS was used to identify read-enriched regions (peaks) with P value < 1e–10. The MAnorm package was used for the quantitative comparison of signals between samples with the following criteria: |M value| > 1 and P < 0.05.
For RNA-seq, featureCount program of the Subread package was used to determine the RNA-seq read density for the genes. The DEseq2 program was used for detecting differentially expressed genes based on the following criteria: |log2 fold-change| > 1 and P < 0.05.
For Bisulphite-seq, the extent of the cytosine methylation was determined with the bismark_methylation_extractor implemented in the Bismark program. Next, the methylation ratio of a cytosine was calculated as the number of mCs divided by the number of reads covering the position. Bases covered by fewer than three reads were considered low-confidence positions whose methylation ratios were not recorded.
Genome_build: IGDBv1.0
Supplementary_files_format_and_content: peak files and bigwig files
Supplementary_files_format_and_content: FPKM values for each sample
 
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)
GSE167228 Evolutionary rewiring of wheat abiotic stress responsive network by lineage-specific transposable elements I
GSE167229 Evolutionary rewiring of wheat abiotic stress responsive network by lineage-specific transposable elements
Relations
BioSample SAMN18026052
SRA SRX10149534

Supplementary data files not provided
SRA Run SelectorHelp
Raw data are available in SRA
Processed data are available on Series record

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