|
| Status |
Public on Aug 11, 2016 |
| Title |
Data Heifer #10991_11v14 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Blasto_heifer_11
|
| Organism |
Bos taurus |
| Characteristics |
sample type: in_vitro
developmental stage: early_blastocyst
breed (cattle): holstein
time of embryo collection: 168 hpf (hours post-in-vitro fertilization)
heifer age: 11 months
tissue or cell type: embryo
|
| Treatment protocol |
Ovarian stimulation: Each heifer was first treated with progesterone (CIDR) during the luteal phase. The dominant follicle was aspirated 36 hours prior to administration of hormones. The ovarian stimulation program consisted of 6 injections of 30mg FSH ad
Embryo production: In vitro maturation (IVM), in vitro fertilization (IVF), in vitro culture (IVC)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from the pools using the AllPrep DNA/RNA micro kit, following the Parallel gDNA and total RNA extraction protocol. Total RNA integrity and concentration were evaluated on an Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto
Amp protocol: To generate enough material for hybridization, the samples were linearly amplified. Antisense RNA (aRNA) was produced using the RiboAmp®HSPlus RNA Amplification kit (Thermo fisher Scientific). After two amplification rounds of 6 hr each, the
|
| Label |
Cy5
|
| Label protocol |
For each sample, aRNA was labeled using the ULS Fluorescent Labeling Kit for Agilent arrays (Cy3/Cy5) (Kreatech Diagnostics, Amsterdam, Netherlands). The labeled product was then purified with the Pico-Pure RNA Isolation Kit.
|
| |
|
| Channel 2 |
| Source name |
Blasto_heifer_14
|
| Organism |
Bos taurus |
| Characteristics |
sample type: in_vitro
developmental stage: early_blastocyst
breed (cattle): holstein
time of embryo collection: 168 hpf (hours post-in-vitro fertilization)
heifer age: 14 months
tissue or cell type: embryo
|
| Treatment protocol |
Ovarian stimulation: Each heifer was first treated with progesterone (CIDR) during the luteal phase. The dominant follicle was aspirated 36 hours prior to administration of hormones. The ovarian stimulation program consisted of 6 injections of 30mg FSH ad
Embryo production: In vitro maturation (IVM), in vitro fertilization (IVF), in vitro culture (IVC)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from the pools using the AllPrep DNA/RNA micro kit, following the Parallel gDNA and total RNA extraction protocol. Total RNA integrity and concentration were evaluated on an Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto
Amp protocol: To generate enough material for hybridization, the samples were linearly amplified. Antisense RNA (aRNA) was produced using the RiboAmp®HSPlus RNA Amplification kit (Thermo fisher Scientific). After two amplification rounds of 6 hr each, the
|
| Label |
Cy3
|
| Label protocol |
For each sample, aRNA was labeled using the ULS Fluorescent Labeling Kit for Agilent arrays (Cy3/Cy5) (Kreatech Diagnostics, Amsterdam, Netherlands). The labeled product was then purified with the Pico-Pure RNA Isolation Kit.
|
| |
|
| |
| Hybridization protocol |
The hybridization of microarray slides 4x44K printed with oligonucleotides by Agilent for all EmbryoGENE related projects using the expression array protocol.
|
| Scan protocol |
Scan on a PowerScanner (Tecan) using the autogain function. Images were quantified using ArrayPro version 6.3 (MediaCybernetics)
|
| Description |
Rep 2 of 4
|
| Data processing |
Background subtracted, Loess and Quantile normalized data obtained from log2 of processed Red signal/processed Green signal. Flexarray software used.
|
| |
|
| Submission date |
Aug 10, 2016 |
| Last update date |
Aug 11, 2016 |
| Contact name |
Léonie Morin-Doré |
| Organization name |
Laval University
|
| Department |
Animal Sciences
|
| Street address |
2440, boulevard Hochelaga
|
| City |
Quebec |
| State/province |
Quebec |
| ZIP/Postal code |
G1V 0A6 |
| Country |
Canada |
| |
|
| Platform ID |
GPL13226 |
| Series (2) |
| GSE85437 |
Transcriptomic evaluation of bovine embryo quality on blastocysts obtained from peri-pubertal oocyte donors [11 mo vs 14 mo] |
| GSE85438 |
Transcriptomic evaluation of bovine embryo quality on blastocysts obtained from peri-pubertal oocyte donors |
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