|
| Status |
Public on Oct 23, 2015 |
| Title |
Tobacco(L2)_normal_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
transgenic tobacco, normal condition, replication 1
|
| Organism |
Nicotiana benthamiana |
| Characteristics |
genotype/variation: CbFAD3 transgenic
condition: normal condition
tissue: leaf
age: two weeks old
|
| Treatment protocol |
Two-week-old seedlings of L2 and WT tobaccos were treated with 200 mM NaCl for 6 h. Leaves from three independent plants of each sample were collected and stored in liquid N2 immediately.
|
| Growth protocol |
WT and transgenic tobacco seeds were sown in soil (30% peat, 70% tuff), and grown at 25℃ under 16 h light/8 h dark.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was prepared using the TRIzol reagent following the manufacturer's recommendations. RNA was quantified using a NanoDrop-1000 spectrophotometer.
|
| Label |
Cy3
|
| Label protocol |
cDNA labeled with a fluorescent dye (Cy3-dCTP) was produced by Eberwine’s linear RNA amplification method and subsequent enzymatic reaction. This procedure has been previously described, and the procedure has been improved by using CapitalBio cRNA Amplification and Labeling Kit (CapitalBio) for producing higher yields of labeled cDNA.
|
| |
|
| Hybridization protocol |
DNA in hybridization solution was denatured at 95℃ for 3 min prior to loading onto a microarray. Arrays were hybridized was preformed in a Agilent Hybridization Oven overnight at a rotation speed of 20 rpm at 42℃ and washed with two consecutive solutions (0.2% SDS, 2× SSC at 42℃ for 5 min, and 0.2× SSC for 5 min at room temperature).
|
| Scan protocol |
After hybridization and post-hybridization washes, slides were scanned immediately in Agilent Microarray Scanner with Surescan High Resolution Technology (Agilent Technologies, Santa Clara, CA).
|
| Description |
Gene expression in transgenic tobaccos under normal condtion
|
| Data processing |
Feature Extraction v10.7.3.1 (Agilent Technologies, CA) software was used to extract all features of the data obtained from the scanned images and genespring software was used to analyze the raw data, which are normalized by percentile normalization.
|
| |
|
| Submission date |
Oct 22, 2015 |
| Last update date |
Oct 23, 2015 |
| Contact name |
Yulan Shi |
| E-mail(s) |
sylbaby15@lzb.ac.cn
|
| Organization name |
Cold and Arid Regions Environmental and Engineering Research Institution
|
| Street address |
Donggang road 320#
|
| City |
Lanzhou |
| ZIP/Postal code |
730000 |
| Country |
China |
| |
|
| Platform ID |
GPL21056 |
| Series (1) |
| GSE74260 |
Gene expression signatures of transgenic and wild-type tobaccos under normal or salt-stress conditions |
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