|
Status |
Public on Jun 12, 2013 |
Title |
51725Pre |
Sample type |
RNA |
|
|
Source name |
skeletal muscle
|
Organism |
Homo sapiens |
Characteristics |
subject: 51725Pre age: 20 time: prior to exercise training tissue: skeletal muscle biopsy
|
Treatment protocol |
Subjects were physiologically assessed at baseline
|
Growth protocol |
Not applicable
|
Extracted molecule |
total RNA |
Extraction protocol |
Paired tissue samples (obtained before and after exercise training) of ~20 mg each were processed simultaneously in 1 mL TRIzol (Invitrogen) using a Mini-Beadbeater-8 (Biospec Inc.) for 15 sec on the homogenize setting. After 5 min of incubation at room temperature, 200 µL of chloroform (Sigma-Aldrich) was added and samples shaken vigorously by hand. Samples were briefly incubated on ice prior to centrifugation at 12,000 g for 15 min. The supernatant was removed and mixed with isopropanol (Sigma-Aldrich) and spun once more at 12,000 g for 10 min, after 10 min of incubation. After a single washing step with 75% EtOH RNA pellets were re-suspended in 40 µL DEPC-treated water (Ambion) and quantified using a NanoDrop Spectrophotometer (NanoDrop Technologies).
|
Label |
biotin
|
Label protocol |
Reverse transcription of RNA was carried out using the Affymetrix 3′ IVT express kit. 100 ng of total RNA was reverse transcribed as per manufacturer's protocol, and quantified using a Nanodrop ND-1000 instrument. aRNA was fragmented and labeled as per manufacturers protocol and hybridized to Affymetrix U133+2 arrays (Affymetrix, USA).
|
|
|
Hybridization protocol |
Arrays were washed, stained and scanned following Affymetrix standard procedures, using an Affymetrix 3000 7G scanner and Affymetrix 450 wash station. A visual inspection of each array was carried out.
|
Scan protocol |
Affymetrix Gene ChIP Scanner 3000 7G
|
Description |
gene expression prior to exercise training HER_110_51725Pre
|
Data processing |
The data were analyzed with Microarray Suite version 5.0 (MAS 5.0) using Affymetrix default analysis settings and global scaling as normalization method.
|
|
|
Submission date |
Jun 12, 2013 |
Last update date |
May 30, 2024 |
Contact name |
James Timmons |
E-mail(s) |
jamie@augurprecisionmedicine.com, jamie.timmons@gmail.com
|
Organization name |
Augur Precision Medicine LTD
|
Street address |
Bridge of Allan
|
City |
Stirling |
ZIP/Postal code |
FK9 4AX |
Country |
United Kingdom |
|
|
Platform ID |
GPL570 |
Series (1) |
GSE47874 |
The Heritage (HEalth, RIsk factors, exercise Training And GEnetics) family study |
|