|
| Status |
Public on May 31, 2020 |
| Title |
MytilusHyedulis18vs22 [12_2] |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Hybrids_18°C
|
| Organism |
Mytilus edulis x Mytilus galloprovincialis |
| Characteristics |
Sex: Mytilus edulis male
temperature: 18°C
|
| Growth protocol |
Microplate with oocytes and sperms were incubated in a temperature-controlled chamber al 18°C; 22°C for 48h in the dark.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted from 200.000 larvae from each experimental condition using Trizol following manufacturer's instructions
|
| Label |
Cy3
|
| Label protocol |
5 µg of total RNA were primed with 0.5 µg oligodT(19)VN primer at 70°C for 10 min, then reversed transcribed at 42°C for 2 h in the presence of 400 U ReverseAid MuLV H minus RTase (Fermentas), and 100 µM each dATP, dTTP, dGTP, with 25 µM dCTP, 25 µM Cy3-labeled dCTP or Cy5-label dCTP
|
| |
|
| Channel 2 |
| Source name |
Hybrids_22°C
|
| Organism |
Mytilus edulis x Mytilus galloprovincialis |
| Characteristics |
Sex: Mytilus edulis male
temperature: 22°C
|
| Growth protocol |
Microplate with oocytes and sperms were incubated in a temperature-controlled chamber al 18°C; 22°C for 48h in the dark.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted from 200.000 larvae from each experimental condition using Trizol following manufacturer's instructions
|
| Label |
Cy5
|
| Label protocol |
5 µg of total RNA were primed with 0.5 µg oligodT(19)VN primer at 70°C for 10 min, then reversed transcribed at 42°C for 2 h in the presence of 400 U ReverseAid MuLV H minus RTase (Fermentas), and 100 µM each dATP, dTTP, dGTP, with 25 µM dCTP, 25 µM Cy3-labeled dCTP or Cy5-label dCTP
|
| |
|
| |
| Hybridization protocol |
43°C, 16/20 h, hybridization buffer Northern Max (Ambion). Pre-hybridization for 1h in same conditions.
|
| Scan protocol |
Scanned on Chip Reader (Applied biosystems Laboratories) at 5 µm resolution
|
| Data processing |
Offset background subtracted, global mean normalisation across microarray surfaces and local mean normalisation across element signal intensity. Data obtained from log2 of processed Ch2 signal / processed Ch1 signal.
|
| |
|
| Submission date |
Mar 26, 2019 |
| Last update date |
Jun 01, 2020 |
| Contact name |
Gerolamo Lanfranchi |
| E-mail(s) |
stefano.cagnin@unipd.it
|
| Phone |
+39-0498276219
|
| Organization name |
University of Padova
|
| Department |
CRIBI - Biotechnology Center and Biology Department
|
| Lab |
Functional Genomics Lab
|
| Street address |
Via U. Bassi, 58/B
|
| City |
Padova |
| ZIP/Postal code |
35131 |
| Country |
Italy |
| |
|
| Platform ID |
GPL22172 |
| Series (1) |
| GSE128849 |
Molecular mechanisms underlying heat stress effects on Mytilus edulis and galloprovincialis hybrids at early larval stages |
|
