|
| Status |
Public on Feb 01, 2006 |
| Title |
C57BL/6J-Ghrhr<lit> (lit/lit) mice injected with growth hormone (GH), replicate 5 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
C57BL/6J-Ghrhr<lit> (lit/lit), purchased from the Jackson Laboratories, Bar Harbor, Maine
|
| Organism |
Mus musculus |
| Characteristics |
two lit/lit mice, age 28 days, bone (including femur and tibia)
|
| Biomaterial provider |
GH provided by Biosidus Co., Buones Aires, Argentina
|
| Treatment protocol |
GH (4mg/kg body weight) injected subcutaneously
mice euthanized at 24 hours after growth hormone injection
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Bone tissues were ground with Trizol (Invitrogen, Carlsbad, CA). Total RNA was then extracted with an RNeasy Mini Kit (Qiagen Inc., Valencia, CA), and DNase treated with a DNA-free kit (Ambion, Austin, TX)
|
| Label |
Cy5
|
| Label protocol |
Total bone RNA was reverse transcribed and linear amplified using a primer containing oligo (dT) and a T7 RNA polymerase promoter. After synthesis of the first and second strands of cDNA, the product was used in an in vitro transcription reaction to generate cRNA in the presence of cyanine 5-labeled UTP
|
| |
|
| Channel 2 |
| Source name |
C57BL/6J-Ghrhr<lit> (lit/lit), provided by the Jackson Laboratories, Bar Harbor, Maine
|
| Organism |
Mus musculus |
| Characteristics |
two lit/lit mice, age 28 days, bone (including femur and tibia)
|
| Biomaterial provider |
PBS were purchased from Sigma
|
| Treatment protocol |
PBS injected subcutaneously
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Bone tissues were ground with Trizol (Invitron, Carlsbad, CA). Total RNA was then extracted with an RNeasy Mini Kit (Qiagen Inc., Valencia, CA), and DNase treated with a DNA-free kit (Ambion, Austin, TX)
|
| Label |
Cy3
|
| Label protocol |
Total bone RNA was reverse transcribed and linear amplified using a primer containing oligo (dT) and a T7 RNA polymerase promoter. After synthesis of the first and second strands of cDNA, the product was used in an in vitro transcription reaction to generate cRNA in the presence of cyanine 3-labeled UTP
|
| |
|
| |
| Hybridization protocol |
Two micrograms of fragmented cyanine 3-labeled cRNA of control sample was mixed with equal amounts of cyanine 5-labeled cRNA of GH treated sample, and the mixtures were incubated for 17 hours at 60°C with constant rotation. The slide was then washed in sodium, saline citrate (SSC) buffer, and dried with pressurized nitrogen
|
| Scan protocol |
All slides were scanned using an Agilent DNA Microarray scanner and the images were processed and analyzed using Agilent feature extraction software
|
| Description |
Output file name: US14702375_16011472013405_S01_A01.txt
|
| Data processing |
Data were analyzed using GeneSpring 7.0 (Silicon Genetics). Local background-subtracted median signal intensities were used as intensity measures and the data were normalized using LOWESS normalization. The genes which had a raw signal of 100 or greater and passed with flag value present were included in the analysis
|
| |
|
| Submission date |
Nov 28, 2005 |
| Last update date |
Nov 29, 2005 |
| Contact name |
Hongrun Yu |
| E-mail(s) |
[email protected]
|
| Phone |
(909) 825-7084
|
| Fax |
(909) 796-1680
|
| Organization name |
J.L. Pettis Memorial VA Medical Center
|
| Department |
Musculoskeletal Disease Center
|
| Street address |
11201 Benton Street (151)
|
| City |
Loma Linda |
| State/province |
CA |
| ZIP/Postal code |
92357 |
| Country |
USA |
| |
|
| Platform ID |
GPL922 |
| Series (1) |
| GSE3689 |
Whole genome microarray analysis of growth hormone induced genes in bone |
|
