|
Status |
Public on Dec 09, 2015 |
Title |
Heart_sedentary_24wk_rep3 |
Sample type |
RNA |
|
|
Source name |
Heart_sedentary_24wk
|
Organism |
Rattus norvegicus |
Characteristics |
strain: Wistar Kyoto age: 24 weeks old Sex: male sample group: sedentary tissue: heart chip batch id: chip 2
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was extracted from frozen heart using the Micro-to-Midi Total RNA Purification System kit with Trizol and DNase on-column digestion (Invitrogen, Melbourne, Australia). RNA integrity was verified and the concentration determined on the Experion Automated Electrophoresis System (Bio-Rad Laboratories, NSW, Australia).
|
Label |
cy3
|
Label protocol |
Microarray gene expression analysis was performed by the Australian Genome Research Facility (AGRF, Melbourne, Australia). The quality and quantity of human total RNA was ascertained on the Agilent Bioanalyser 2100 using the NanoChip protocol. A total of 500 ng of total RNA was then prepared for hybridisation to the Ilumina RatRef-12_V1_0_R4
|
|
|
Hybridization protocol |
A total hybridisation volume of 15ul was prepared for each sample and 15ul loaded into a single array on the Ilumina RatRef-12 BeadChip. The chip was then hybridised at 58C for 16 hours in an oven with a rocking platform. Following hybridisation, the chip was washed using the appropriate protocols as outlined in the Illumina manual.
|
Scan protocol |
Upon completion of the washing, the chips were then coupled with Cy3 and scanned in the Illumina iScan Reader. The scanner operating software, BeadStudio, converts the signal on the array into a TXT file for analysis.
|
Description |
group3 rat 3
|
Data processing |
The sample probe profile and control probe profile files were read into R (v3.2.2), and underwent normexp background correction with limma (v3.24.15) using negative control probes and quantile normalization using negative and positive control probes.
|
|
|
Submission date |
Dec 08, 2015 |
Last update date |
Dec 09, 2015 |
Contact name |
Glenn David Wadley |
E-mail(s) |
glenn.wadley@deakin.edu.au
|
Organization name |
Deakin University
|
Street address |
221 Burwood Highway
|
City |
Burwood |
State/province |
Victoria |
ZIP/Postal code |
3125 |
Country |
Australia |
|
|
Platform ID |
GPL6101 |
Series (1) |
GSE75781 |
Endurance training in early life results in long-term programming of heart mass in rats |
|