|
| Status |
Public on Jan 01, 2015 |
| Title |
wild-type E14,5 DRG replicate3 - technical replicate 2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
E14,5 DRG extract from wild type embryos
|
| Organism |
Mus musculus |
| Characteristics |
age: embryonic day 14,5 (E14,5)
tissue: dorsal root ganglion (DRG)
genotype/variation: wild type
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Spinal columns from E14 embryos from multiple litters were quickly dissected in Hank's Buffered Salt Solution (HBSS) and placed in RNAlater (Ambion) overnight at 4°C awaiting genotyping. DRGs were then dissected out and homogenized using glass beads in Precellys 24 (Bertin Technologies, France), followed by RNA extraction using RNeasy mini kit (Qiagen). RNA was then concentrated by precipitation with lithium chloride (2.5M final concentration). The amount and quality of RNA was assessed using NanoDrop (NanoDrop Technologies) and Experion system (BIO-RAD). Three independent replicates, each containing DRGs from 6-8 embryos from multiple litters, were prepared for each genotype.
|
| Label |
Cy5
|
| Label protocol |
For the preparation of the labelled Cy3- and Cy5- aRNA target, one microgram of total RNA samples were amplified and labelled using Amino Allyl Message Amp II aRNA amplification kit (Ambion, Austin, TX, USA), according the manufacturer’s instructions.
|
| |
|
| Channel 2 |
| Source name |
Total RNA from pooled mouse E14,5 and adult DRGs
|
| Organism |
Mus musculus |
| Characteristics |
tissue: dorsal root ganglion (DRG)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Spinal columns from E14 embryos from multiple litters were quickly dissected in Hank's Buffered Salt Solution (HBSS) and placed in RNAlater (Ambion) overnight at 4°C awaiting genotyping. DRGs were then dissected out and homogenized using glass beads in Precellys 24 (Bertin Technologies, France), followed by RNA extraction using RNeasy mini kit (Qiagen). RNA was then concentrated by precipitation with lithium chloride (2.5M final concentration). The amount and quality of RNA was assessed using NanoDrop (NanoDrop Technologies) and Experion system (BIO-RAD). Three independent replicates, each containing DRGs from 6-8 embryos from multiple litters, were prepared for each genotype.
|
| Label |
Cy3
|
| Label protocol |
For the preparation of the labelled Cy3- and Cy5- aRNA target, one microgram of total RNA samples were amplified and labelled using Amino Allyl Message Amp II aRNA amplification kit (Ambion, Austin, TX, USA), according the manufacturer’s instructions.
|
| |
|
| |
| Hybridization protocol |
The two labelled aRNA were mixed, fragmented and hybridized on one array of a Nimblegen 12-plex microarray according the manufacturer's instructions.
|
| Scan protocol |
Scanned on an Innopsys Innoscan900 scanner
|
| Description |
Biological replicate 3 of 3. Technical replicate 2. E14,5 DRG extract from wild type embryos.
|
| Data processing |
Images were quantified using Nimblescan V2.5. The rawdata were normalized by Loess (Limma Package).
|
| |
|
| Submission date |
Jan 09, 2013 |
| Last update date |
Jan 01, 2015 |
| Contact name |
Aziz Moqrich |
| E-mail(s) |
aziz.moqrich@univ-amu.fr
|
| Organization name |
CNRS
|
| Street address |
case 907, campus de luminy
|
| City |
Marseille |
| ZIP/Postal code |
13009 |
| Country |
France |
| |
|
| Platform ID |
GPL15902 |
| Series (1) |
| GSE43391 |
Murine E14,5 DRGs: TrkAC-KI vs. Wild type |
|
