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| Status |
Public on Apr 19, 2010 |
| Title |
RNA-Seq on libraries made from serial dilutions of Drosophila melanogaster S2 cell mRNA and ERCC external RNA controls |
| Organisms |
Drosophila melanogaster; synthetic construct |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
RNA-Seq on libraries made from serial dilutions of mRNA from Drosophila melanogaster S2 cell and the External RNA Controls Consortium (ERCC) external RNA controls.
We evaluated performance of RNA-Seq by serially diluting a complex pool of known synthetic PolyA+ mRNAs from the External RNA Controls Consortium (ERCC) and PolyA+ mRNA from Drosophila S2 cells. ERCC mRNAs were obtained under Phase V testing from the National Institutes of Standards and Technology (NIST). The ERCC pool contained 96 species of mRNA of various lengths and GC content covering a 2^20 concentration range. Libraries were constructed with 100ng to 10pg of input mRNA. Our data shows an outstanding linear fit between RNA-Seq read density and known input amounts.
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| Overall design |
We performed RNA-Seq from libraries made with 0.01ng to 100ng mRNA (mixture of mRNA from Drosophila melanogaster S2 cells and ERCC RNA controls). One RNA-Seq library was prepared with 100ng mRNA and six libraries were made with serial dilutions of mRNA using a modified protocol (see Sample 'extraction protocol'). One lane was sequenced for each library and all seven libraries were run on the same flow cell.
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| Contributor(s) |
Zhang Y, Jiang L, Oliver B |
| Citation(s) |
21816910 |
| |
| Submission date |
Feb 27, 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 |
| |
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| Platforms (1) |
| GPL10119 |
Illumina Genome Analyzer II (Drosophila melanogaster; synthetic construct) |
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| Samples (7)
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| GSM516588 |
100ng_library_methA_S2_2.5%ERCC_phaseV_pool15mRNA |
| GSM516589 |
100ng_library_methB_S2_2.5%ERCC_phaseV_pool15_mRNA |
| GSM516590 |
50ng_library_methB_S2_2.5%ERCC_phaseV_pool15_mRNA |
| GSM516591 |
10ng_library_methB_S2_2.5%ERCC_phaseV_pool15_mRNA |
| GSM516592 |
1ng_library_methB_S2_2.5%ERCC_phaseV_pool15_mRNA |
| GSM516593 |
0.4ng_library_methB_S2_2.5%ERCC_phaseV_pool15_mRNA |
| GSM516594 |
0.01ng_library_methB_S2_2.5%ERCC_phaseV_pool15_mRNA |
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| Relations |
| SRA |
SRP002255 |
| BioProject |
PRJNA125199 |
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