|
| Status |
Public on Dec 01, 2006 |
| Title |
Gastrocnemius tissue at time 0 from young mouse, biological rep5 |
| Sample type |
RNA |
| |
|
| Source name |
gastrocnemius tissue in control-fed 5 month-old C57BL/6 mice
|
| Organism |
Mus musculus |
| Characteristics |
Age: 5 month-old
Diet: normal
|
| Treatment protocol |
Mice were fed one of two life-long dietary regimens: normal diet (ND) and caloric restriction (CR). Mice on ND were provided acidic water ad libitum and fed 84 kcal/wk which is ~15% less than the average ad libitum diet. C57BL/6NHsd mice on this dietary regiment typically have an average lifespan of 30 mo. in our colony. Mice on CR were fed a diet that resulted in a 26% difference in caloric intake to the ND animals. The diet fed to CR mice was enriched in protein, vitamins and minerals such that CR and ND mice were fed nearly identical amounts of these components.
|
| Growth protocol |
Male C57BL/6NHsd mice were purchased from Harlan Sprague-Dawley at 6-7 weeks of age and individually housed for the remainder of their lifespan.
|
| Extracted molecule |
polyA RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions and polyadenylate [poly(A)+] RNA was purified from the total RNA with oligo-dT-linked Oligotex resin (Qiagen, Valencia, CA).
|
| Label |
biotin
|
| Label protocol |
One microgram of poly(A)+ RNA was converted into double-stranded cDNA (ds-cDNA) by using the SuperScript Choice System (Life Technologies) with an oligo-dT primer containing a T7 RNA polymerase promoter (Genset, La Jolla, CA). The ds-cDNA was extracted with phenol-chloroform-isoamyl alcohol and precipitated with pellet paint coprecipitant (Novagen, Madison, WI). Biotin-labeled RNA was synthesized in vitro using a high-yield RNA transcript labeling kit (BioArray; Enzo, Farmingdale, NY). The biotin-labeled antisense cRNA was purified using the RNeasy affinity column (Qiagen) and fragmented randomly.
|
| |
|
| Hybridization protocol |
The hybridization cocktail (200 µl) containing 10 µg of fragmented cRNA was injected into the MOE430A array (Affymetrix, Santa Clara, CA). The GeneChip was placed in a 45°C oven at 60 rpm for 16 h. After hybridization, the GeneChips were washed and stained in a fluidic station (model 800101, Affymetrix) with signal amplification protocol using antibody.
|
| Scan protocol |
GeneChips were scanned at a resolution of 3 µm twice using a Hewlett-Packard GeneArray Scanner (model 900154, Affymetrix), and the averaged images were used for further analysis.
|
| Description |
Gene expression data from gastrocnemius tissue from 5 month-old mice on normal diet
gastrocnemius tissue at time 0 from young mouse, biological rep5
|
| Data processing |
Robust Multi-Array Analysis (RMA) was used to pre-process and normalize the raw Affymetrix GeneChip data. Microarray Suite version 5.0 (MAS5.0) was used to compute absent/present calls.
|
| |
|
| Submission date |
Nov 20, 2006 |
| Last update date |
Nov 21, 2006 |
| Contact name |
Michael Edwards |
| E-mail(s) |
[email protected]
|
| Phone |
303-724-6054
|
| Organization name |
UC Denver
|
| Department |
Pulmonary
|
| Street address |
12700 East 19th Avenue, Box C272
|
| City |
Aurora |
| State/province |
CO |
| ZIP/Postal code |
80045 |
| Country |
USA |
| |
|
| Platform ID |
GPL339 |
| Series (1) |
| GSE6323 |
Expression data from skeletal muscle of young, old and old calorie restricted mice |
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