|
| Status |
Public on Oct 14, 2009 |
| Title |
Horse #3 T0 vs T1 Rep1 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Equine skeletal muscle at T0
|
| Organism |
Equus caballus |
| Characteristics |
exercise protocol: Skeletal muscle biopsy from gluteus medius of an unconditioned 4 year old Thoroughbred gelding pre exercise
tissue: gluteus medius
age: 4 year old
gender: gelding
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was amplified using a MessageAmpTM amplified RNA (aRNA) linear amplification kit (Ambion).
|
| Label |
Alexa Fluor 555
|
| Label protocol |
aRNA was reverse transcribed and directly labelled with Flour647 or Flour555 using the SuperScript™ Plus Direct cDNA Labeling System (BD Biosciences)
|
| |
|
| Channel 2 |
| Source name |
Equine skeletal muscle at T1
|
| Organism |
Equus caballus |
| Characteristics |
exercise protocol: Skeletal muscle biopsy from gluteus medius of an unconditioned 4 year old Thoroughbred gelding immediately (<8 minutes) post exercise
tissue: gluteus medius
age: 4 year old
gender: gelding
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was amplified using a MessageAmpTM amplified RNA (aRNA) linear amplification kit (Ambion).
|
| Label |
Alexa Fluor 647
|
| Label protocol |
aRNA was reverse transcribed and directly labelled with Flour647 or Flour555 using the SuperScript™ Plus Direct cDNA Labeling System (BD Biosciences)
|
| |
|
| |
| Hybridization protocol |
Microarray hybridisations were performed on an automated HS400 hybridisation station (Tecan Ltd.) with the following protocol - Wash: 75°C, runs 1, wash 10 s, soak 20 s; probe injection: 85°C; denaturation: 95°C, 2 min; hybridisation: 42°C, agitation frequency medium, 4hrs; hybridisation: 35°C, agitation frequency medium, 4hrs; hybridisation: 30°C, agitation frequency medium, 4hrs; wash: 37°C, runs 2, wash 10 s, soak 20 s; wash: 25°C, runs 2, wash 15 s, soak 30 s; wash: 25°C, runs 2, wash 20 s, soak 40 s; slide drying: 25°C 2mins.
|
| Scan protocol |
Hybridised and dried slides were scanned with a GenePix 4000B scanner (Axon Instruments Inc.) using the GenePix 6.0 microarray image analysis package (Axon Instruments Inc.).
|
| Description |
Unconditioned thoroughbred geldings were exercised to maximal heart rate or fatigue on an equine high-speed treadmill. Skeletal muscle biopsies were taken from the middle gluteal muscle before, immediately after and four hours after exercise.
|
| Data processing |
Background correction (Limma “rma”) and print tip lowess normalization were performed. Log(base2) ratios were calculated for each spot on the microarray and duplicate spots were averaged.
|
| |
|
| Submission date |
May 26, 2009 |
| Last update date |
Oct 14, 2009 |
| Contact name |
Beatrice Ann McGivney |
| E-mail(s) |
beatrice.mcgivney@ucd.ie
|
| Phone |
352872852916
|
| Organization name |
University College Dublin
|
| Department |
School of Agriculture, Food Science and Veterinary Medicine
|
| Lab |
Transcriptomics Laboratory
|
| Street address |
Belfield
|
| City |
Dublin |
| State/province |
Dublin |
| ZIP/Postal code |
4 |
| Country |
Ireland |
| |
|
| Platform ID |
GPL8582 |
| Series (1) |
| GSE16235 |
Equine skeletal muscle: pre versus post-exercise |
|
