|
| Status |
Public on Nov 15, 2013 |
| Title |
lumbar motor neurons bio rep 3 |
| Sample type |
RNA |
| |
|
| Source name |
motor neurons laser captured from sections of L5 levesl of the spinal cord of P7 male mouse
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6J
gender: male
age: postnatal day 7
cell type: lumbar motor neurons
|
| Treatment protocol |
none
|
| Growth protocol |
Mice were obtained from Jackson Labs and were housed under standerd condicition as per IACUC protocol.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Postnatal day 7 male animals were perfused with 30% sucrose, lumbosacral spinal cord and midbrain regions were rapidly recovered, embedded in OCT compound, and frozen in liquid nitrogen. 12 µm-thick cryosections were mounted on RNAse-free, PEN-foil covered glass slides (Zeiss), fixed for 2 min in 100% EtOH, rinsed in 50% EtOH, stained with 1% cresyl violet for 2 min, rinsed with 50% EtOH, dehydrated in graded solutions of ethanol and air dried prior to LCM using PALM Microbeam system. From each animal, ~200 DL, L5, and oculomotor motor neurons were collected directly in lysis buffer. RNA was purified using Absolutely RNA, NanoPrep kit. RNA integrity was assessed on the Bioanalyzer 2100.
|
| Label |
biotin
|
| Label protocol |
At least 1.5 ng of purified RNA was the starting material used in the WT-Ovation Pico RNA Amplification System (Nugen, San Carlos,CA) with the FL-Ovation cDNA Biotin Module V2 (Nugen) to generate labeled probe.
|
| |
|
| Hybridization protocol |
µg of biotinylated cRNA from three independent samples for each motor neuron group isolated by LCM was hybridized to on Affymetrix Mouse Genome 430 2.0 Arrays.
|
| Scan protocol |
Arrays were scanned on an Affymetrix Scanner 3000-7G scanner with GCOS software. Scanning was peformed according to the protocol described in the Affymetrix GeneChip® Expression Analysis Technical Manual, November 2004 Edition
|
| Description |
gene expression data from lumbar motor neurons of young wildtype mouse
|
| Data processing |
GeneSpring and BioconductorLIMMA package was used for statistical analysis
|
| |
|
| Submission date |
Nov 05, 2013 |
| Last update date |
Nov 15, 2013 |
| Contact name |
Krista Joan Spiller |
| E-mail(s) |
[email protected]
|
| Organization name |
Columbia University
|
| Street address |
630 W 168th street
|
| City |
New York |
| State/province |
New York |
| ZIP/Postal code |
10032 |
| Country |
USA |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE52118 |
Comparison of gene expression in motor pools with differential vulnerability in ALS |
|
