|
| Status |
Public on Dec 20, 2012 |
| Title |
J LF brain rep2 |
| Sample type |
RNA |
| |
|
| Source name |
mutant mouse brain 2 days low-fat diet
|
| Organism |
Mus musculus |
| Characteristics |
strain: BALB/cByJ (Acads-/-)
genotype: Acads-/-
tissue: brain
treatment: low-fat diet
|
| Treatment protocol |
After weaning the offspring were treated the same; from weaning until 12 weeks of age mice were fed a chow diet (LabDiets 5001) ad libitum. Several days prior to initiation of experimental diets, bedding was removed and replaced with stainless steel wire floor inserts. Nesting tubes were provided to reduce time spent on wire flooring. At 12 weeks of age mice were fed ad libitum a high-fat diet D12331 or low-fat diet D12329 (Research Diets) for 2 days.
|
| Growth protocol |
Twelve-week old male mice were singly housed in filter-top cages and kept under 12h light/12h dark conditions in a specific-pathogen free facility fed a Chow diet (LabDiet 5001).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Tissues were quickly removed and frozen in liquid nitrogen and stored at -80C for subsequent preparation of total RNA. Total RNA was isolated using TRI-Reagent (Molecular Research Center Inc. Cincinnati, Ohio) with modifications to remove DNA using the Qiagen RNAeasy columns and DNaseI Kit (Qiagen, Valencia, CA). RNA was stored at –70C in RNase-free H2O supplemented with the RNase inhibitor Superasin (Ambion, Austin, TX) as per manufacturers directions. Quality and quantity of RNA was determined using a Agilent Bioanalyzer as per manufacturers procedures (Agilent Technologies, Palo Alto, CA).
|
| Label |
Dig-UTP
|
| Label protocol |
1 ug of total RNA was used to transcribe DIG-labeled cRNA using Applied Biosystems Chemiluminescent RT-IVT Kit V2.0.
|
| |
|
| Hybridization protocol |
Microarray hybridization of each sample (using ten micrograms of fragmented, DIG-labeled cRNA) and processing were performed according to Applied Biosystems protocols.
|
| Scan protocol |
Chemiluminescence detection, imaging, auto gridding, and image analysis was done according to Applied Biosystems protocols and the 1700 Chemiluminescent Microarray Analyzer Software v. 1.0.3.
|
| Data processing |
Signal intensities across microarrays were normalized using the quantile-quantile method (www.bioconductor.org).
|
| |
|
| Submission date |
Jan 18, 2012 |
| Last update date |
Dec 20, 2012 |
| Contact name |
Brenda Richards |
| E-mail(s) |
richarbk@pbrc.edu
|
| Organization name |
Pennington Biomedical Research Center
|
| Department |
Basic Science
|
| Lab |
Genetics of Eating Behavior
|
| Street address |
6400 Perkins Rd.
|
| City |
Baton Rouge |
| State/province |
LA |
| ZIP/Postal code |
70808 |
| Country |
USA |
| |
|
| Platform ID |
GPL2995 |
| Series (1) |
| GSE35180 |
Effects of diet and Acads genotype on transcriptional response in brain and liver |
|
