NCBI Logo
GEO Logo
   NCBI > GEO > Accession DisplayHelp Not logged in | LoginHelp
GEO help: Mouse over screen elements for information.
          Go
Sample GSM6727478 Query DataSets for GSM6727478
Status Public on Sep 26, 2023
Title Cold-H3K9ac
Sample type SRA
 
Source name leaf,H3K9ac,Cold
Organism Oryza sativa Japonica Group
Characteristics tissue: leaf
antibody: Millipore,07-352
treatment: After a 24 h grown at 4℃ for cold treatment(cold)
Treatment protocol At the 13th day, a half of PCR plates with seedlings were continued to grow in the normal conditions (CK) and another half were grown at 4℃ for cold treatment. After a 24 h cold treatment (COLD), two biological replicates of CK and cold treated (COLD) seedlings were collected and stored at −80°C for later use.
Growth protocol Seeds of Nipponbare (Oryza sativa ssp. japonica) were pre-germinated at room temperature under dark conditions for 3 days. Uniformly germinated seeds were individually put into 96-well PCR plates with bottom cut. Each plate with germinated seeds were put into the liquid culture medium for continuing to grow under a mean temperature as 28/22°C and 12 h/12 h for a light-dark cycle in the greenhouse.
Extracted molecule genomic DNA
Extraction protocol Uniformly germinated seeds were individually put into 96-well PCR plates with bottom cut. Each plate with germinated seeds were put into the liquid culture medium for continuing to grow under a mean temperature as 28/22°C and 12 h/12 h for a light-dark cycle in the greenhouse.
Total RNA was extracted from CK and COLD seedlings, respectively, using the RNeasy plant Mini kit (Qiagen, 74904). After removal of genomic DNA contamination using DNase I treatment, approximately 30 µg total RNA was used for mRNA enrichment using oligo (dT) conjugated magnetic beads.The DNA-RNA hybrid immunoprecipitation (DRIP) experiment and DRIP-seq libraries were conducted as described before
 
Library strategy ChIP-Seq
Library source genomic
Library selection ChIP
Instrument model Illumina HiSeq 2500
 
Data processing For RNA-seq analyses, the quality of raw reads was assessed using FastQC (Kroll et al. 2014). Raw reads were cleaned after removal of adapters and poor-quality bases using Trimmomatic (version 0.39) (Bolger et al. 2014). Clean reads were aligned to the rice reference genome using Hisat2 (version 2.1.0) (Kim et al. 2015). The number of reads mapped to each gene was counted using FeatureCounts (version 1.6.4) (Liao et al. 2014). FPKM (Fragments Per Kilobase of exon model per Million mapped fragments) values were calculated to represent the gene expression levels.
For DRIP-seq data analyses, the raw data were processed for removal of adapter and low-quality bases using TrimGalore (version0.4.3,Babraham Bioinformatics). Clean reads were aligned to the rice reference genome (MSU version7.0) using Bowtie (Langmead et al. 2009) with the following parameters‐‐best‐‐strata -m 1 - x 800. Additional duplicate reads were removed by using the SAMtools rmdup,version1.4 (Li et al. 2009). The strand-specific reads were separated using flags of SAM files. The direction of R-loops was determined according to the direction of the R2 reads (R1 for the paired-end reads) aligned to the genome.
Assembly: MSU v7.0
 
Submission date Nov 12, 2022
Last update date Sep 26, 2023
Contact name Zexue He
E-mail(s) hezexue@stu.njau.edu.cn
Organization name Nanjing Agricultural University
Department Agricultural College
Street address Xuanwu District,1 Weigang
City Nanjing
State/province Jiangsu Province
ZIP/Postal code 210018
Country China
 
Platform ID GPL18525
Series (1)
GSE217841 R-loops act as a master regulatory switch modulating transcription of cold-responsive genes in rice
Relations
BioSample SAMN31700878
SRA SRX18243030

Supplementary file Size Download File type/resource
GSM6727478_Cold-H3K9ac.bw 55.3 Mb (ftp)(http) BW
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

| NLM | NIH | GEO Help | Disclaimer | Accessibility |
NCBI Home NCBI Search NCBI SiteMap