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Sample GSM2521697 Query DataSets for GSM2521697
Status Public on Jun 06, 2017
Title S2 T0 H3 ChIP rep1 6.25U MNase
Sample type SRA
 
Source name UPR-induced S2 cells, T0 H3 ChIP 6.25U MNase
Organism Drosophila melanogaster
Characteristics cell type: S2 cell
time: T0 (0 hours, control)
mnase concentration: 6.25
chip antibody: H3 (abcam, ab1791)
Extracted molecule genomic DNA
Extraction protocol [MNase-seq] MNase titration was performed as described previously (Mieczkowski, Cook, Bowman et al. Nat. Communications 2016). For h-MACC, after addition of EDTA/EGTA and SDS half of the digests were kept at 4C as input, the other half were adjusted to ChIP buffer conditions [10mM TRIS ph8.0, 100mM NaCl, 1mM EDTA, 0.1% sodiumdeoxycholate, 0.5% sarkosyl, 1% triton x-100 and complete protease inhibitors (Roche)] with 1ml of ChIP buffer. After tumbling for 10min at 4C, the digests were spun at high speed for 10min at 4C and the supernatant was incubated with anti H3 antibodies (abcam ab1791) as described (Bowman et al. BMC Genomics 2013).
 
Library strategy MNase-Seq
Library source genomic
Library selection MNase
Instrument model Illumina HiSeq 2000
 
Data processing MNase-seq, ATAC-seq and ChIP-seq: The sequenced paired-end reads were mapped to dm3 genome using Bowtie aligner v. 0.12.939. Only uniquely mapped reads with no more than two mismatches were retained. The reads with the insert sizes less than 50 bp or larger than 500 bp were filtered out. Genomic positions with the numbers of mapped tags above the significance threshold of z-score = 7 were identified as anomalous, and the tags mapped to such positions were discarded. Read frequencies were computed in 300-bp non-overlapping bins in the case of fly data and in 500-bp bins in the case of human data for each titration point independently. The read frequencies were normalized by the corresponding library sizes to represent values per one million of mapped reads. To facilitate the comparison of the results between different genomes, the frequencies were additionally scaled by the factors representing ratios between the corresponding genome size and 100 Mb.
RNA-seq: Tags were aligned using Tophat software package with default parameters. RNA-Seq tag frequencies were normalized for GC-content using bioconductor package EDASeq and then the expression estimates for each gene were obtained using bioconductor package DESeq.
Genome_build: dm3
Supplementary_files_format_and_content: bedGraph files representing coverage profiles normalized to library sizes
 
Submission date Mar 04, 2017
Last update date May 15, 2019
Contact name Michael Tolstorukov
E-mail(s) tolstorukov@molbio.mgh.harvard.edu
Organization name Massachusetts General Hospital
Department Molecular Biology
Street address 185 Cambridge Street
City Boston
State/province MA
ZIP/Postal code 02114
Country USA
 
Platform ID GPL13304
Series (1)
GSE95689 Widespread changes in nucleosome accessibility without changes in nucleosome occupancy during a rapid transcriptional induction
Relations
BioSample SAMN06477995
SRA SRX2612471

Supplementary file Size Download File type/resource
GSM2521697_s2.mnase_6.25U_T0.H3.r1_bin200.bedGraph.gz 4.9 Mb (ftp)(http) BEDGRAPH
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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