|
| Status |
Public on Aug 01, 2018 |
| Title |
Spleens of BALB/c mouse, 14th day, control 1 |
| Sample type |
RNA |
| |
|
| Source name |
Spleens,14th day, control
|
| Organism |
Mus musculus |
| Characteristics |
gender: female
age: six week
sample type: control
|
| Treatment protocol |
Treatment groups of mice were injected intraperitoneally with OVA at 2 mg/kg body weight for 3 times (days 1, 3, and 5).The control groups were treated with the solvent vehicle (normal saline).
|
| Growth protocol |
Six-week-old female BALB/c mice were injected intraperitoneally with OVA or normal saline at 2 mg/kg body weight for 3 times (days 1, 3, and 5). At the end of the experiment the animals were humanely sacrificed by pentobarbital anesthetization 2 hours at 14th and 28th day, separately.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA containing small RNA was extracted from the snap-frozen spleens by using the Trizol reagent (Invitrogen, Carlsbad, CA, USA) and purified with mirVana miRNA Isolation Kit (Ambion, Austin, TX, USA) according to manufacturer’s protocol.
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 0.5 ug RNA using the One-Color Low RNA Input Linear Amplification PLUS kit (Agilent) according to the manufacturer's instructions, followed by RNAeasy column purification (QIAGEN, Valencia, CA). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
|
| |
|
| Hybridization protocol |
1.5 ug of Cy3-labelled cRNA (specific activity >10.0 pmol Cy3/ug cRNA) was fragmented at 60°C for 30 minutes in a reaction volume of 250 ml containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 250 ml of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to LncRNA+mRNA Mouse Gene Expression Microarray V1.0 for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent), then dried immediately by brief centrifugation.
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2505B) using one color scan setting for 1x44k array slides (Scan Area 61x21.6 mm, Scan resolution 10um, Dye channel is set to Green and Green PMT is set to 100%).
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 9.1 (Agilent) using default parameters (protocol GE1-v1_91 and Grid: 012391_D_20060331) to obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded.
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| |
|
| Submission date |
Jul 30, 2018 |
| Last update date |
Aug 01, 2018 |
| Contact name |
LI Yong Ning |
| E-mail(s) |
liyongning@cfsa.net.cn
|
| Phone |
13466796251
|
| Organization name |
China National Center for Food Safety Risk Assessment
|
| Street address |
7 Panjiayuan Nanli
|
| City |
Beijing |
| ZIP/Postal code |
100021 |
| Country |
China |
| |
|
| Platform ID |
GPL22782 |
| Series (2) |
| GSE117900 |
Expression data from OVA-induced allergic mice [Agilent] |
| GSE117916 |
Expression data from OVA-induced allergic mice |
|
