|
| Status |
Public on Mar 06, 2012 |
| Title |
Stretch 1 hour sample 8 |
| Sample type |
RNA |
| |
|
| Source name |
Stretch 1 hour
|
| Organism |
Rattus norvegicus |
| Characteristics |
cell type: Primary alveolar epithelial cell
strain: Sprague Dawley
gender: male
weight: 200-300g
|
| Treatment protocol |
Isolated type II cells from whole lung, grown for 5 days in MEM + 0.1x FBS until cells attain type I characteristics
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Quiagen miRNAeasy Kit per protocol; quality control with Agilent 2100 bioanalyzer
|
| Label |
Hy5
|
| Label protocol |
Adjust RNA and volume to 0.25ug to 1ug in 2ul of nuclease free water; add CIP buffer, enzyme, and spike-in RNA at 37C x 30min; quench 95C x 5min; snap cool on ice 2-15min; mix with Labeling buffer, enzyme, dye, and DMSO 16C x 1hr; quench 65C x 15min, cool to 4C; hybridization within 1 hour
|
| |
|
| Hybridization protocol |
Mix dyed samples with warmed hyb buffer; denature 95C x 3min; snap cool on ice 2-15min; centrifuge max 1min; apply sample to Agilent coverslip and slide, and assemble hyb apparatus; hyb 56C x 16hr rotation speed 4; wash 3x with buffers A, B, C 2min each, rinse with water, dry by centriguge 800rpm x 3min
|
| Scan protocol |
Images acquired at 635 and 532 nm using GenePix 4000B, analyzed genePix pro 6.0.1.27
|
| Description |
Each array was hybridized with each sample labeled with Hy5 against a Common Reference (labeled Hy3). Common Reference is an equal (same ng of RNA) mix of all sample groupings (Control, 1hr stretch, 6hr stretch) from each of the 12 individual samples pooled together.
|
| Data processing |
The software was Partek Genomics Suite (version 6.4) where background subtracted intensities were extracted, any background intensity below 1 was changed to 1, and loess normalization was applied.
|
| |
|
| Submission date |
Mar 04, 2012 |
| Last update date |
Mar 06, 2012 |
| Contact name |
Nadir Yehya |
| E-mail(s) |
yehyan@email.chop.edu
|
| Organization name |
University of Pennsylvania
|
| Department |
Bioengineering
|
| Street address |
390 Towne Bldg, U Penn
|
| City |
Philadelphia |
| State/province |
PA |
| ZIP/Postal code |
19104 |
| Country |
USA |
| |
|
| Platform ID |
GPL7722 |
| Series (1) |
| GSE36256 |
Differential expression of miRNA in stretched rat alveolar epithelial cells |
|
