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Status |
Public on Aug 20, 2013 |
Title |
neonatal rat beta cells: endocrine cells from postnatal day 2-3 pancreas of male and female rats, bio rep 1 |
Sample type |
RNA |
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Source name |
neonatal rat beta cells: endocrine cells, FACS-sorted for their high side scatter (HSSC), from postnatal day 2-3 pancreas of male and female Wistar rats, and containing approximately 65-70% insulin+ cells
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Organism |
Rattus norvegicus |
Characteristics |
cell type: neonatal rat beta cells gender: male and female strain: Wistar age: 2-3 d
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Treatment protocol |
None, freshly isolated beta cells snap frozen in liquid nitrogen immediately after isolation
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Growth protocol |
Pancreatic beta cells were FACS-purified from neonatal (postnatal day 2-3)and 10-weeks old Wistar rat pancreas, and immediately snap frozen in liquid nitrogen, prior to total RNA extraction in batch, from all biological replicates.
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Extracted molecule |
total RNA |
Extraction protocol |
RNEasy extraction of total RNA was performed according to the manufacturer's instructions. RNA integrity verified on Bioanalyzer with minimal RIN cutoff = 8.
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Label |
biotin
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Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 2 ug total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
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Hybridization protocol |
After fragmentation at 94°C for 35 min in fragmentation buffer (40mM Tris, 30mM magnesium acetate, 10mM potassium acetate), the labelled cRNA was hybridized for 16h to the Affymetrix HG133A
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Scan protocol |
Arrays were stained with phycoerythrin-streptavidin with the Affymetrix Fluidics Station 400, and scanned in the Affymetrix GeneArray2500scanner.
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Description |
neonatal rat beta cells: endocrine cells, FACS-sorted for their high side scatter (HSSC), from postnatal day 2-3 pancreas of male and female Wistar rats, and containing approximately 65-70% insulin+ cells, biological replicate 1, pool of 30 animals
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Data processing |
Scanned arrays were analyzed with dChip model-based expression analysis to the array with the median intensity and correction for mismatch hybridization using the perfect match-mismatch correction option.
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Submission date |
May 22, 2013 |
Last update date |
Aug 20, 2013 |
Contact name |
Geert A. Martens |
E-mail(s) |
geert.martens@vub.ac.be
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Organization name |
Vrije Universiteit Brussel
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Department |
Diabetes Research Center
|
Street address |
Laarbeeklaan 103
|
City |
Brussels |
ZIP/Postal code |
1090 |
Country |
Belgium |
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Platform ID |
GPL1355 |
Series (1) |
GSE47174 |
Genome-wide mRNA expression profiles of FACS-enriched rat beta cells freshly isolated from neonatal (postnatal day 2-3) and adult (10 weeks-old) Wistar rats |
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