|
Status |
Public on Jul 16, 2009 |
Title |
Retina C57BL/6J 2 days rep2 |
Sample type |
RNA |
|
|
Source name |
retina, C57BL/6J, 2 days post crush
|
Organism |
Mus musculus |
Characteristics |
strain: C57BL/6J age: 60-90 days gender: Male tissue: retina from both eyes
|
Treatment protocol |
Optic nerve crush: Mice were deeply anesthetized with a mixture of 13 mg/kg of Rompum and 87 mg/kg of Ketalar. A small incision was made in the lateral aspect of the conjunctiva. With a pair of small forceps the edge of the conjunctiva next to the globe was retracted slightly and rotated laterally, allowing visualization of the posterior aspect of the globe where the optic nerve could be observed. Viewed under a binocular operating microscope, the surrounding connective tissue and muscle was gently separated from the nerve. The exposed optic nerve was grasped for 10 sec with a pair of Dumont cross-clamp #7 forceps (Roboz, cat. #RS=5027, Gaithersburg, MD). This instrument was chosen because its spring action applied a moderate yet constant and consistent force to the optic nerve. The forceps were then removed and the eye was allowed to rotate back into place.
|
Extracted molecule |
total RNA |
Extraction protocol |
The tissue was homogenized and extracted according to the RNA-Stat-60 protocol as described by the manufacturer (Tel-Test, Friendswood, TX). The quality and purity of RNA was assessed using an Agilent Bioanalyzer 2100 system.
|
Label |
Cy3
|
Label protocol |
The RNA from each sample was processed with the Illumina TotalPrep RNA Amplification Kit (Ambion, Austin, TX) to produce labeled cRNA.
|
|
|
Hybridization protocol |
Standard Protocol for Illumina Sentrix 6 Mouse BeadChip microarrays
|
Scan protocol |
Illumina BeadArray Reader and BeadScan scan protocol
|
Description |
C57BL/6J retina, total RNA isolation 2 days after optic nerve crush: The exposed optic nerve was grasped for 10 sec with a pair of Dumont cross-clamp #7 forceps
|
Data processing |
Using Microsoft Excel, we normalized the data using the formula 2 (z-score of log2 [intensity]) + 8
|
|
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Submission date |
Jul 15, 2009 |
Last update date |
Jul 15, 2009 |
Contact name |
William Edwin Orr |
E-mail(s) |
weorr@utmem.edu
|
Phone |
901 448-3635
|
Fax |
901 448-5028
|
Organization name |
University of Tennessee Health Science Center
|
Department |
Ophthalmology
|
Lab |
Vision Core
|
Street address |
930 Madison Avenue, Suite 757
|
City |
Memphis |
State/province |
TN |
ZIP/Postal code |
38163 |
Country |
USA |
|
|
Platform ID |
GPL4234 |
Series (1) |
GSE17117 |
Differential response of C57BL/6J mouse and DBA/2J mouse to optic nerve crush |
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