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Sample GSM1000400 Query DataSets for GSM1000400
Status Public on Jan 17, 2013
Title S2 STARR-seq biological replicate 1
Sample type SRA
 
Source name S2 cells
Organism Drosophila melanogaster
Characteristics cell type: S2 cells
sample type: PCR amplified cDNA (STARR-seq transcript)
Extracted molecule polyA RNA
Extraction protocol Genomic DNA (source: embryos of the sequenced strain: y; cn bw sp) was isolated, sheared and size selected (~500bp). Following the instructions of NEBNext® DNA Library Prep Reagent Set for Illumina® (NEB; cat. no. E6000L), Illumina Multiplexing Adapters (Illumina Inc; cat. no. PE-400-1001) were ligated and homology arms for In-Fusion® recombination were added by PCR, followed by recombination into the STARR-seq vector (pGL3-Promotor backbone (Promega; cat. no. E1751) with the sequence between BglII and FseI replaced with the following sequence, containing a Drosophila Synthetic Core Promoter (DSCP) (1), an ORF (sgGFP, Qbiogene, Inc), a ccdB suicide gene flanked by homology arms (used for cloning the genomic enhancer candidates during library generation), and the pGL3s SV40 late polyA-signal.). The In-Fusion® reactions were transformed (MegaX DH10B; Invitrogen), grown in liquid culture and plasmids were isolated.
 
Library strategy OTHER
Library source transcriptomic
Library selection cDNA
Instrument model Illumina Genome Analyzer IIx
 
Description PCR amplified cDNA (STARR-seq transcript)
Data processing Basecall where performed using Real-Time Analysis (RTA) version > 1.12.4.2 or CASAVA 1.9.1
Reads were mapped to the dm3 genome using bowtie (bowtie -p 12 -f -X 2000 -v 3 -m 1 --best --strata --quiet INDEX -1 reads_1.fa -2 reads_2.fa) excluding chrU, chrUextra, and chrM. For all subsequently analysis we used only reads collapsed on chromosome, start, end, strand (fragments). Significantly enriched regions (peaks) were called with an in-house pipeline using read density profiles of cDNA and input and an hypergeometric test to assign a p-value to each peak. Enrichment values were corrected within a 95% confidence interval taking into account the number of independent fragments at a single peak summit position.
Genome_build: dm3
Supplementary_files_format_and_content: All processed data files are in plain text. For STARR-seq we report for each peak the chromosome, the summit position, the enrichment over input (log2-scale) at the summit, and the p-value.
 
Submission date Sep 10, 2012
Last update date May 15, 2019
Contact name Daniel Gerlach
E-mail(s) [email protected]
Organization name Boehringer Ingelheim RCV GmbH & Co KG
Department Global Computational Biology and Digital Sciences
Street address Dr.-Boehringer-Gasse 5-11
City Vienna
ZIP/Postal code 1121
Country Austria
 
Platform ID GPL11203
Series (1)
GSE40739 Genome-wide quantitative enhancer activity maps identified by STARR-seq
Relations
Reanalyzed by GSM1397685
SRA SRX187073
BioSample SAMN01176858

Supplementary file Size Download File type/resource
GSM1000400_S2_STARRseq_rep1.peaks.txt.gz 36.4 Kb (ftp)(http) TXT
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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