|
| Status |
Public on Jun 09, 2007 |
| Title |
Pancreas at e12.5, Chip B, rep 2 |
| Sample type |
RNA |
| |
|
| Source name |
Pancreas at e12.5
|
| Organism |
Mus musculus |
| Characteristics |
Affymetrix GeneChip Murine Genome U74 Version 2 Set MG-U74B
|
| Growth protocol |
Time mated NMRI mice (M&B, Ry, Denmark) were killed by cervical dislocation at embryonic day 12.5, 13.5, 14.5, 15.5, or 16.5. Embryos were placed in ice cold PBS and decapitated, and the dorsal pancreas was removed by dissection with DUMONT #SS Biologie forceps (Fine Science Tools) using an inverted microscope. Only dorsal pancreatic buds were isolated due to the difficulty of isolating ventral pancreas at early stages. Dorsal pancreata from a litter were pooled and stored in RNAlater (Ambion) at 4C until RNA isolation (no more than one week).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Pancreata stored in RNAlater were collected by centrifugation at 100 x g for 15 minutes followed by removal of RNAlater. The tissue was homogenized in Eppendorf tubes by addition of 500μl TRIzol (GibcoBRL/Life Technologies) and grinding with disposable pistils (Buch & Holm, Denmark). Total RNA isolation was performed as described in the standard TRIzol procedure. Following RNA isolation genomic DNA contamination was eliminated using DNAfree (Ambion) according to manufacturer’s instructions and total RNA was purified on RNeasy columns (Qiagen) following the standard protocol. RNA integrity was assessed by gel electrophoresis using 1% ethidium bromide containing agarose gels. The amount of isolated RNA was quantified spectrophotometrically on an UltroSpec 2100pro (Amersham Bioscience).
|
| Label |
biotin
|
| Label protocol |
Standard Affymetrix labelling protocol was followed.
|
| |
|
| Hybridization protocol |
Standard Affymetrix hybridization protocol was followed.
|
| Scan protocol |
Standard Affymetrix hybridization protocol was followed.
|
| Description |
Gene expression data from dorsal pancreas at e12.5, MG-U74B version2
|
| Data processing |
The data were analyzed using the R environment and Bioconductor packages and expression summaries were based on the default RMA algorithm.
|
| |
|
| Submission date |
Jun 08, 2007 |
| Last update date |
Jun 08, 2007 |
| Contact name |
Andreas Petri |
| E-mail(s) |
andp@novonordisk.com
|
| Organization name |
Novo Nordisk A/S
|
| Department |
Molecualr Genetics
|
| Street address |
Novo Nordisk Park D6 2.154
|
| City |
Maaloev |
| ZIP/Postal code |
2760 |
| Country |
Denmark |
| |
|
| Platform ID |
GPL82 |
| Series (1) |
| GSE8070 |
Expression profiling of pancreas development |
|
