|
| Status |
Public on Jan 01, 2014 |
| Title |
RNA from brain of stimulated silkworm_P2 |
| Sample type |
RNA |
| |
|
| Source name |
brain, stimulated, replicate2
|
| Organism |
Bombyx mori |
| Characteristics |
tissue: brain
gender: male
treatment protocol: female-odor stimulated
|
| Treatment protocol |
Adult male silkworm moth were stimulated with the female odor, which has sex pheromone components, for 30 min. The whole brains were collected by disecction.
|
| Growth protocol |
The lavae of silkworms were fed with artificial diet (silkmate). The egg and larvae were raised at 25 degree and 12L/12D condition.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The genomic DNA was removed with TurboDNA-free kit (Life Technologies Corporation, Carlsbad, CA). RNA was quantified using a NanoDrop-1000 spectrophotometer and quality was monitored with the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA).
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 0.4 ug RNA using the Low RNA Input Fluorescent Linear Amplification 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 |
1650 ng 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 50 ul containing Agilent fragmentation buffer and Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 55 ul of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent Silkworm Custom EST Microarray 4x44K 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 by raise it slowly from buffer 2.
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2565BA) using one color scan setting for 4x44k array slides (Scan Area 61x21.6 mm, Scan resolution 5um, Dye channel is set to Green. Scan mode is eXtended Dynamic range Scan mode and PMT is set to 10% and 100%).
|
| Description |
Gene Expression in brain of female-odor stimulated male silkworm
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 9.1.3.1 (Agilent) using default parameters (protocol GE1-v5_91_0806 and Grid: 014879_D_20070820) to obtain background subtracted and spatially detrended Processed Signal intensities.
|
| |
|
| Submission date |
Jul 12, 2012 |
| Last update date |
Jan 01, 2014 |
| Contact name |
Takumi Nishiuchi |
| E-mail(s) |
tnish9@staff.kanazawa-u.ac.jp
|
| Organization name |
Kanazawa University
|
| Street address |
13-1 Takaramachi
|
| City |
Kanazawa |
| ZIP/Postal code |
920-0934 |
| Country |
Japan |
| |
|
| Platform ID |
GPL15776 |
| Series (1) |
| GSE39306 |
Identification of Hr38 as a neural activity-induced gene |
|
