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Status |
Public on Apr 19, 2010 |
Title |
100ng_S2_2.5ng_ERCC_phaseV_pool15_mRNA-seq |
Sample type |
SRA |
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|
Source name |
Drosophila S2 cell and 2.5ng ERCC RNA controls
|
Organisms |
Drosophila melanogaster; synthetic construct |
Characteristics |
cell line: S2 external rna control: ERCC phase V pool 15 s2 treatment: msl2 RNAi rna quantity (s2): 100ng rna quantity (ercc): 2.5ng
|
Treatment protocol |
S2 cells were mock or RNAi treated with 10ng dsRNA using standard procedure. See Zhang et al., PLoS Biology 2010 for details.
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Growth protocol |
Drosophila S2 cells ("SL2" originally obtained from the Drosophila RNAi Screening Center, Boston MA) were grown at 25°C in Schneider's Drosophila Medium (Invitrogen, Carlsbad, CA) supplemented with 10% Fetal Bovine serum (SAFC Biosciences, Lenexa, KS) and Penicillin-Streptomycin.
|
Extracted molecule |
total RNA |
Extraction protocol |
Libraries were made with 100ng S2 mRNA or 100ng mRNA from S2 RNAi-treated cells with 5ng, 2.5ng or 1ng ERCC mRNAs and with 50ng ERCC mRNAs only following the standard procedure. See Zhang et al. PLoS Biology 2010 for details.
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Library strategy |
RNA-Seq |
Library source |
transcriptomic |
Library selection |
cDNA |
Instrument model |
Illumina Genome Analyzer II |
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Description |
Sequences and abundance information of ERCC phase V pool 15 used in this study were provided separately with the supplementary files: ERCC_phaseV_pool15_sequence.txt ERCC_phaseV_pool15_abundance.txt
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Data processing |
We retained reads that passed default quality filtering parameters in the Illumina pipeline (v1.4.0). We aligned reads passing quality filtering using Bowtie (v0.10.0; parameter -m 1 -v 2). Sequences of the ERCC RNAs (*ERCC_map.txt) were used as the alignment reference. Additionally, Drosophila genome sequence (BDGP release 5, dm3) and ERCC RNA sequences were combined into a reference (*ERCC_dm3_map.txt). We used unique mapped reads with less than 2 mismatches for analysis.
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Submission date |
Mar 01, 2010 |
Last update date |
May 15, 2019 |
Contact name |
Brian Oliver |
E-mail(s) |
briano@nih.gov
|
Phone |
301-204-9463
|
Organization name |
NIDDK, NIH
|
Department |
LBG
|
Lab |
Developmental Genomics
|
Street address |
50 South Drive
|
City |
Bethesda |
State/province |
MD |
ZIP/Postal code |
20892 |
Country |
USA |
|
|
Platform ID |
GPL10119 |
Series (1) |
GSE20579 |
RNA-Seq on libraries made from ERCC external RNA controls, and a mixture of D. melanogaster S2 cell mRNA and ERCC mRNAs |
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Relations |
Reanalyzed by |
GSM3274609 |
SRA |
SRX019235 |
BioSample |
SAMN00011329 |