Identification of E93 binding sites in Drosophila melanogaster
Ontology highlight
ABSTRACT:
PROVIDER: PRJEB11893 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| ERR1146213.fastq.gz | Fastqsanger.gz | |||
| ERR1146214.fastq.gz | Fastqsanger.gz |
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Project description:Chromatin immunoprecipitation was performed with modification of the protocols described before (Lee et al., 2006; Lilja et al., 2007). Briefly, the chorion was removed from 12-hour old embryos, cross-linked using 2% paraformaldehyde and resuspended in storage buffer (50mM Tris-HCl pH8.0, 1 mM EDTA). Embryos were then lysed in SDS-lysis buffer (1.0 ml 5 M NaCl, 2.5 ml 1 M Tris-HCl pH 8.0, 0.5 ml 0.5M EDTA, 2.5 ml 10 % SDS), resuspended in SDS-lysis and Triton buffer (1.0 ml 5 M NaCl, 5.0 ml 1 M Tris-HCl pH 8.0, 0.5 ml 0.5M EDTA, 2.5 ml Triton X-100, protease inhibitor cocktail) and sonicated on ice to an average length of 350 bp. The resulting sheared chromatin (25Izg) was subjected to immunoprecipitation using anti-Myc antibody (Santa Cruz) as described previously (Lee et al., 2006). ChIP-sequencing libraries were constructed following manufactureras instructions (Illumina). The resulting DNA libraries were sequenced using Illumina platform (University of Massachusetts Medical School Core Facility).
2019-02-02 | E-MTAB-4083 | biostudies-arrayexpress
Project description:Whole-genome analysis of heat shock factor binding sites in Drosophila melanogaster. Heat shock factor IP DNA or Mock IP DNA from heat shocked Kc 167 cells compared to whole cell extract on Agilent 2x244k tiling arrays.
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Project description:Whole-genome analysis of heat shock factor binding sites in Drosophila melanogaster using Heat Shock factor IP or Mock IP vs Whole cell Extract on Agilent 2x244k tiling arrays Heat Shock Factor IP or MOCK IP vs Whole Cell Extract
2011-01-05 | E-GEOD-19744 | biostudies-arrayexpress
Project description:The 3' ends of most Drosophila melanogaster genes are poorly annotated or are determined by only a single EST or cDNA clone. To enhance the annotation of poly(A) site use in Drosophila, we performed deep sequencing on RNA isolated from 29 dissected tissues using an approach designed to enrich for poly(A) spanning reads. From these experiments, we identified 1.4 million poly(A) spanning reads leading to the identification of many new poly(A) sites and the identification of many tissue-specific poly(A) sites. For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf
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Project description:The 3' ends of most Drosophila melanogaster genes are poorly annotated or are determined by only a single EST or cDNA clone. To enhance the annotation of poly(A) site use in Drosophila, we performed deep sequencing on RNA isolated from 29 dissected tissues using an approach designed to enrich for poly(A) spanning reads. From these experiments, we identified 1.4 million poly(A) spanning reads leading to the identification of many new poly(A) sites and the identification of many tissue-specific poly(A) sites. For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf RNA from 29 dissected Drosophila melanogaster tissues (in duplicate) were used to prepare polyA enriched RNA-Seq libraries. Briefly, total RNA was poly(A) selected, fragmented, and ligated to 5' and 3' RNA linkers. These libraries were amplified using Illumina paired-end primers, and subsequently reamplified using a 3' primer complementary to the 3' adapter but containing 6 Ts at the 3' end. The libraries were also multiplexed and up to 12 samples mixed per lane and sequenced on an Illumina GAIIx using paired-end 76 bp reads, or an illumina HiSeq 2000 using paired-end 100 bp reads. All reads were mapped to the Drosophila melanogaster genome to identify unmapped reads. Unmapped reads containing at least 10 A residues at the 3' end were identified, the terminal A residues trimmed, realigned to the genome to identify uniquely mapped reads. Such reads were identified as polyA spanning reads
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Project description:Identification of Esg genomic binding sites in Drosophila testes
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Project description:How temporal cues combine with spatial inputs to control gene expression during development is poorly understood. Here, we test the hypothesis that the Drosophila transcription factor E93 controls temporal gene expression by regulating chromatin accessibility. Precocious expression of E93 early in wing development reveals that it can simultaneously activate and deactivate different target enhancers. Notably, the precocious patterns of enhancer activity resemble the wild-type patterns that occur later in development, suggesting that provision of E93 alters the competence of enhancers to respond to spatial cues. Genomic profiling reveals that precocious E93 expression is sufficient to regulate chromatin accessibility at a subset of its targets. These accessibility changes mimic those that normally occur later in development, indicating that precocious E93 accelerates the wild-type developmental program. Further, we find that target enhancers that do not respond to precocious E93 in early wings become responsive after a developmental transition, suggesting that parallel temporal pathways work alongside E93. These findings support a model wherein E93 expression functions as an instructive cue that defines a broad window of developmental time through control of chromatin accessibility.
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Project description:Analysis of Drosophila melanogaster nascent transcription start sites
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