|
| Status |
Public on Mar 18, 2010 |
| Title |
protoplasts at T1, biological rep2 [CPK] |
| Sample type |
RNA |
| |
|
| Source name |
Arabidopsis leaves
|
| Organism |
Arabidopsis thaliana |
| Characteristics |
tissue: mesophyll protoplasts
genotype: Col-0
age: 28 days, leaf 5-7
protocol: expression of constitutively active CPK5ac
|
| Treatment protocol |
Protoplasts (0.8 - 2 million) were transfected with 1 mg DNA/ml protoplasts using PEG-calcium and incubated in 10 ml 0.5 M mannitol, 4 mM MES pH 5.7 and 20 mM KCl for 6 hr at 25oC.
|
| Growth protocol |
Arabidopsis plants (Col-0) were grown in a controlled room for 28 days at 22oC, 60% relative humidity, 75 uE light and 12 hr photoperiod.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using RNeasy plant mini kit (Qiagen) according to the manufacturer’s protocol
|
| Label |
biotin
|
| Label protocol |
Total RNA (10 µg) were amplified, biotinylated and fragmented using the BioArrayTM HighYieldTM RNA transcript labelling kit (Enzo) according to the manufacturer’s instructions.
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized on the Arabidopsis ATH1 GeneChips using the Affymetrix 640 Hybridization Oven. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using GeneChip Scanner 7G with AutoLoader.
|
| Description |
Gene expression data from Arabidopsis mesophyll protoplasts transfected with CPK5ac DNA for 6hr
|
| Data processing |
The data were analyzed in FlexArray with RMA including background correction and normalization.
|
| |
|
| Submission date |
Jun 06, 2009 |
| Last update date |
Mar 12, 2010 |
| Contact name |
Jen Sheen |
| E-mail(s) |
[email protected]
|
| Organization name |
Massachusetts General Hospital
|
| Department |
Molecular Biology
|
| Lab |
Sheen
|
| Street address |
185 Cambridge st. CPZN7620
|
| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02114-2790 |
| Country |
USA |
| |
|
| Platform ID |
GPL198 |
| Series (2) |
| GSE16471 |
Identification of CPK5ac and CPK11ac early target genes in Arabidopsis mesophyll cells |
| GSE16557 |
Gene expression changes underlying initial cellular and molecular responses to MAMPs in Arabidopsis mesophyll cells |
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