|
| Status |
Public on Mar 15, 2014 |
| Title |
SAMPLE 12: TW ED |
| Sample type |
RNA |
| |
|
| Source name |
Theca wall EarlyDom
|
| Organism |
Equus caballus |
| Characteristics |
cell type: theca wall
developmental stage: EarlyDom
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA from granulosa and theca cells was extracted using TRIzol® according to manufacturer’s instructions (Life Technologies, Paisley, UK) and analysed using the Agilent Bioanalyzer 2100 (Agilent Technologies).
|
| Label |
Cy3
|
| Label protocol |
RNA of good quality (RIN>7.0) was reverse-transcribed (500 ng) with an oligo (dT) primer bearing a T7 promoter followed by cDNA purification and in vitro transcription with T7 polymerase to generate antisense RNA (aRNA) using the MessageAmp aRNA Kit (Life Technologies), as per manufacturer’s instructions. This was then purified, analysed on the Agilent Bioanalyzer 2100 and used to generate Cy3-labelled aRNA
|
| |
|
| Hybridization protocol |
Cy3-labelled aRNA was fragmented at 60C for 30 min and hybridised onto the Agilent Horse Gene Expression Microarray (version 2010; Agilent Technologies, Wokingham, UK) at 65C for 17 h using Agilent hybridisation chambers and oven.
|
| Scan protocol |
Slides were washed and scanned with an Axon scanner (Molecular Devices LLC, Sunnyvale, CA, USA).
|
| Description |
Gene expression in theca wall of early dominant follicles
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 9.1 (Agilent) using default parameters to obtain background subtracted and spatially detrended Processed Signal intensities. Foreground log2(intensities) were normalized by quantile normalization .
|
| |
|
| Submission date |
Nov 05, 2013 |
| Last update date |
Mar 15, 2014 |
| Contact name |
Xavier Donadeu |
| E-mail(s) |
xavier.donadeu@roslin.ed.ac.uk
|
| Organization name |
Roslin Institute, University of Edinburgh
|
| Street address |
Easter Bush
|
| City |
Midlothian |
| ZIP/Postal code |
EH25 9RG |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL15189 |
| Series (1) |
| GSE52109 |
Transcriptome profiling of dominant follicle development in the equine ovary |
|
