|
| Status |
Public on Apr 29, 2009 |
| Title |
after 3 days in mannitol medium rep2 |
| Sample type |
RNA |
| |
|
| Source name |
100904A_DH46_4d2
|
| Organism |
Hordeum vulgare |
| Characteristics |
after 3 days in mannitol medium
|
| Treatment protocol |
Anthers extracted from the spikes under a stereoscopic microscope were inoculated in a treatment medium containing 0.7 M mannitol, 40 mM CaCl2, and 8 g/l agarose, and kept at 25 C in the dark for 4 days (Cistue´et al. 2003). Samples were collected before and after 4 days in mannitol medium. Three samples from each step were harvested and used for microarray analysis.
|
| Growth protocol |
Donor plants were grown in growth chambers with controlled temperature (18-21ºC), relative humidity (70-80%) and photoperiod (16 h light), as described by Cistué et al. (2003).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using TRIzol reagent (Invitrogen) from frozen crown samples taken from above soil tissue excluding most fully extended lamina. The protocol is available on the Arabidopsis Information Resource website at http://www.arabidopsis.org/info/2010_projects/comp_proj/AFGC/RevisedAFGC/site2RnaL.htm. RNA was purified using Qiagen RNeasy spin columns with DNase treatment. Quality was assessed by running 25-250 ng on a RNA Lab-On-A-Chip (Caliper Technologies) using an Agilent Bioanalyzer 2100 (Agilent Technologies).
|
| Label |
biotin
|
| Label protocol |
The BioArray High-Yield RNA Transcript Labeling Kit (Enzo Diagnostics) was then used to synthesize biotin-labeled cRNA from template cDNA by in vitro transcription.
|
| |
|
| Hybridization protocol |
10 µg labeled, fragmented cRNA was hybridized at 45°C with rotation for 16 hours in an Affymetrix GeneChip Hybridization Oven 320 on Affymetrix Barley1 Genechip arrays. The arrays were washed and stained using streptavidin phycoerythrin on an Affymetrix Fluidics Station 400.
|
| Scan protocol |
The arrays were scanned on a Hewlett-Packard GeneArray scanner.
|
| Description |
after 3 days in mannitol medium
|
| Data processing |
GCOS MAS 5
|
| |
|
| Submission date |
Jan 30, 2008 |
| Last update date |
Apr 28, 2009 |
| Contact name |
Livia Tommasini |
| E-mail(s) |
liviat@ucr.edu
|
| Phone |
951 827 3808
|
| Fax |
951 827 4437
|
| URL |
http://plantbiology.ucr.edu/research_areas/?genomics
|
| Organization name |
University of California, Riverside
|
| Department |
Botany and Plant Science
|
| Lab |
Tim Close
|
| Street address |
2150 Batchelor Hall
|
| City |
Riverside |
| State/province |
CA |
| ZIP/Postal code |
92521-0124 |
| Country |
USA |
| |
|
| Platform ID |
GPL1340 |
| Series (1) |
| GSE10330 |
Transcriptome analysis of barley anthers: effect of mannitol treatment on microspore embryogenesis |
|
