|
| Status |
Public on Sep 30, 2014 |
| Title |
Control 4 |
| Sample type |
RNA |
| |
|
| Source name |
Amygdala tissue, Home Cage
|
| Organism |
Mus musculus |
| Characteristics |
treatment: control
tissue: Brain Amygdala
gender: male
age: adult
genetic background: C57/Bl6J
|
| Treatment protocol |
Adult male mice were either left in their home cage (Control) or subjected to auditory fear conditioning (Fear) using 5 Tone+Shock pairings (6kHz tone, 0.6mA shock).
|
| Growth protocol |
N/A
|
| Extracted molecule |
total RNA |
| Extraction protocol |
30 minutes after conditioning, Fear30 animals were rapidly decapitated and brains fresh frozen on dry ice. The same was done for the Control group. Brains were punched on a freezing microtome using a 1mm punch, and bilateral amygdala tissue were collected from each animal. miRNA and RNA were extracted from the brain punches using the Qiagen miRNeasy Kit.
|
| Label |
Hy3
|
| Label protocol |
A reference pool of RNA was made from mixing equal amount of total RNA from the individual animals. 500 ng of total RNA from each sample and reference pool were labelled with miRCURY™ Hy3™ and Hy5™ fluorescent label (Exiqon, Denmark). The Hy3™-labeled samples and a Hy5™-labeled reference pool RNA samples were mixed pair-wise and hybridized to the miRCURY™ LNA array (6th gen - hsa, mmu & rno) (Exiqon, Denmark) according to manufacturer's protocol for Maui hybridisation chamber.
|
| |
|
| Hybridization protocol |
https://www.exiqon.com/ls/Documents/Scientific/miRCURY-LNA-microRNA-Array-6th-gen-hsa-mmu-rno-manual.pdf (see manufacturer's protocol for more detail)
|
| Scan protocol |
Images were acquired using an Agilent array scanner and spot signals were processed (default background subtraction) and quantified using GenePix Pro 4.1.
|
| Data processing |
Intensities within arrays were normalized using the global Lowess (LOcally WEighted Scatterplot Smoothing) regression algorithm as implemented in the Comprehensive R-based Microarray Analysis web front end (CARMAweb 1.4). The log2 values of the difference between the sample and reference signals were used for hierarchical clustering.
|
| |
|
| Submission date |
Jul 03, 2014 |
| Last update date |
Sep 30, 2014 |
| Contact name |
Brian George Dias |
| Organization name |
Emory University
|
| Department |
Psychiatry and Behavioral Sciences
|
| Street address |
954 Gatewood Rd NE
|
| City |
Atlanta |
| State/province |
GA |
| ZIP/Postal code |
30324 |
| Country |
USA |
| |
|
| Platform ID |
GPL11434 |
| Series (1) |
| GSE59072 |
miRNA and consolidation of cued fear memory |
|
