|
| Status |
Public on Mar 15, 2014 |
| Title |
S2-G2-5 |
| Sample type |
RNA |
| |
|
| Source name |
S2 cells, G2
|
| Organism |
Drosophila melanogaster |
| Characteristics |
cell cycle stage: G2
replicate: 5
cell type: S2 cells
|
| Treatment protocol |
Wing discs and S2 cells were subject to cytometric analysis and partitioned into G1 and G2/M populations. Robust gating was applied based on 2C and 4C peak positions.
|
| Growth protocol |
Drosophila larvae were raised at 25°C at low density until the 3rd instar wandering stage, washed and dissected in PBS (pH 7.4). For S2 cell FACS, low-passage cells (fewer than 30 passages) were used to count and check for viability using a Vi-Cell XR Cell Viability Analyzer (Beckman Coulter) before experiments.
|
| Extracted molecule |
polyA RNA |
| Extraction protocol |
Cells were sorted directly into RNAprotect Cell Reagent (QIAGEN) and total RNA was extracted using QIAGEN RNeasy Plus Micro Kit according to manufacturer instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA was prepared from 100 ng Total RNA using MessageAmp III aRNA Kit from Ambion.
|
| |
|
| Hybridization protocol |
Following fragmentation, 5 ug of cRNA were hybridized for 16 hr at 45C on GeneChip Drosophila Genome 2.0 Arrays. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
GeneChips were scanned using the Hewlett-Packard GeneArray Scanner G2500A.
|
| Data processing |
Array intensities were processed and normalized using the RMA algorithm in the affy library (version affy_1.36.1) from bioconductor in R version 2.15.2.
|
| |
|
| Submission date |
Feb 12, 2014 |
| Last update date |
Mar 15, 2014 |
| Contact name |
Chris W Seidel |
| E-mail(s) |
[email protected]
|
| Phone |
816 926 9054
|
| Organization name |
Stowers Institute
|
| Department |
Genomics
|
| Lab |
Seidel
|
| Street address |
1000 E 50th St
|
| City |
Kansas City |
| State/province |
MO |
| ZIP/Postal code |
64110 |
| Country |
USA |
| |
|
| Platform ID |
GPL1322 |
| Series (2) |
| GSE54926 |
Functional genomic analysis of the periodic transcriptome in the developing Drosophila wing [Affymetrix] |
| GSE54928 |
Functional genomic analysis of the periodic transcriptome in the developing Drosophila wing. |
|
