|
| Status |
Public on Dec 31, 2015 |
| Title |
Follicles female 4_2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Follicles female 4_control
|
| Organism |
Salmo trutta |
| Characteristics |
tissue: Preovulatory follicles
|
| Treatment protocol |
Preovulatory follicles were isolated and incubated in the absence or presence of salmon LH for 18 h at 15 ºC.
|
| Growth protocol |
Preovulatory brown trout (Salmo trutta) females were raised in a local goverment hatchery (Piscifactoria de Bagà, Barcelona, Spain), staged and sacrificed according to approved local animal welfare regulations.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA from brown trout preovulatory follicles was isolated with TRIzol (Invitrogen, Spain) according to the manufacturer’s instructions. The RNA samples were incubated for 20 min at 37° with 10 units of DNaseI (Roche Applied Science) to remove residual genomic DNA prior to purification using the RNeasy MinElute Cleanup kit (Qiagen) according to the RNA clean up protocol. The integrity of the RNA was confirmed by Lab-on-chip analysis using the 2100 Bioanalyzer (Agilent Technologies).
|
| Label |
Cy5
|
| Label protocol |
Amplified and labeled according to Agilent Two-Color Microarray-Based Gene Expression Analysis protocol
|
| |
|
| Channel 2 |
| Source name |
Follicles female 4_LH
|
| Organism |
Salmo trutta |
| Characteristics |
tissue: Preovulatory follicles
|
| Treatment protocol |
Preovulatory follicles were isolated and incubated in the absence or presence of salmon LH for 18 h at 15 ºC.
|
| Growth protocol |
Preovulatory brown trout (Salmo trutta) females were raised in a local goverment hatchery (Piscifactoria de Bagà, Barcelona, Spain), staged and sacrificed according to approved local animal welfare regulations.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA from brown trout preovulatory follicles was isolated with TRIzol (Invitrogen, Spain) according to the manufacturer’s instructions. The RNA samples were incubated for 20 min at 37° with 10 units of DNaseI (Roche Applied Science) to remove residual genomic DNA prior to purification using the RNeasy MinElute Cleanup kit (Qiagen) according to the RNA clean up protocol. The integrity of the RNA was confirmed by Lab-on-chip analysis using the 2100 Bioanalyzer (Agilent Technologies).
|
| Label |
Cy3
|
| Label protocol |
Amplified and labeled according to Agilent Two-Color Microarray-Based Gene Expression Analysis protocol
|
| |
|
| |
| Hybridization protocol |
Overnight hybridization (17 hrs, 65ºC and 10 rpm rotation) was performed in a Microrarray Hybridization Oven (Agilent Technologies). After hybridization, microrarrays were washed with Gene Expression Wash Buffers 1 and 2 (Agilent Technologies).
|
| Scan protocol |
Microrarrays were scanned using the High-Resolution Scanner G2505C (Agilent Technologies).
|
| Description |
Biological replicate 4_C5Cy3
|
| Data processing |
Feature Extraction Software 10.7.3.1 (Agilent Technologies) was used for spot to grid alignment, feature extraction and quantification. Processed data were subsequently imported into GeneSpring GX 11.5 (Agilent Technologies).
|
| |
|
| Submission date |
Apr 06, 2015 |
| Last update date |
Dec 31, 2015 |
| Contact name |
Josep V Planas |
| E-mail(s) |
[email protected]
|
| Phone |
+34-93-4039384
|
| Organization name |
Universitat de Barcelona
|
| Department |
Fisiologia i Immunologia (Biologia)
|
| Lab |
Fisiologia Molecular de Peixos
|
| Street address |
Av. Diagonal 643
|
| City |
Barcelona |
| ZIP/Postal code |
08028 |
| Country |
Spain |
| |
|
| Platform ID |
GPL16819 |
| Series (1) |
| GSE67592 |
Luteinizing hormone induces ovulation via tumor necrosis factor α-dependent increases in prostaglandin F2α in a nonmammalian vertebrate |
|
