|
| Status |
Public on Apr 20, 2007 |
| Title |
Laser capture microdissected (LCM) endosperm at the heart stage, biological replicate 1 |
| Sample type |
RNA |
| |
|
| Source name |
Soybean heart stage endosperm captured by LCM
|
| Organism |
Glycine max |
| Characteristics |
Soybean cv. Asgrow A3237
|
| Treatment protocol |
Pods with lengths between 1.3 and 2.0 cm containing seeds with heart-stage embryos were fixed in 3:1(v/v) ethanol:acetic acid and embedded in paraffin (Kerk et al., Plant Physiol.132. 27-35 (2003)). Seed compartment was captured from eight-micron sections using a Leica AS LMD System.
|
| Growth protocol |
Soybean cv. Asgrow A3237 was grown in the UCLA Plant Growth Center using a 16:8 light-dark cycle at 22°C.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using the PicoPure RNA Extraction Kit (Molecular Devices, Sunnyvale, CA).
|
| Label |
biotin
|
| Label protocol |
Two nanograms of total RNA was amplified using the RiboAmp OA Amplification Kit (Molecular Devices, Sunnyvale, CA). Biotinylated cRNAs were synthesized with the ENZO BioArray High Yield RNA Transcript Labeling Kit using all of the amplified RNA from the RiboAmp OA Kit.
|
| |
|
| Hybridization protocol |
Following fragmentation, ten micrograms of cRNA were hybridized for 16 hr at 45°C with the GeneChip Soybean Genome Array. GeneChips were washed and stained using the EukGE-WS2V4_450 protocol in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner 3000 7G System.
|
| Description |
Endosperm regions from heart stage seeds were microdissected using the LEICA AS LMD system.
|
| Data processing |
The data were analyzed with the Affymetrix GeneChip Operating System 1.3 (GCOS 1.3) using default analysis settings and global scaling as a normalization method. The trimmed mean target intensity of each array was set to 500.
|
| |
|
| Submission date |
Apr 12, 2007 |
| Last update date |
Apr 13, 2007 |
| Contact name |
Bob Goldberg |
| E-mail(s) |
bobglab@mcdb.ucla.edu
|
| Phone |
310-825-3270
|
| Organization name |
University of California, Los Angeles
|
| Department |
Molecular, Cell and Developmental Biology
|
| Street address |
610 Charles E Young Drive East
|
| City |
Los Angeles |
| State/province |
CA |
| ZIP/Postal code |
90095 |
| Country |
USA |
| |
|
| Platform ID |
GPL4592 |
| Series (1) |
| GSE7511 |
Expression data from soybean seed compartments with embryos at the heart stage |
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