|
| Status |
Public on May 31, 2018 |
| Title |
Whole Blood 16hr after 1Gy dose rep 3 |
| Sample type |
RNA |
| |
|
| Source name |
Whole Blood 16hr after 1Gy dose
|
| Organism |
Mus musculus |
| Characteristics |
strain/background: C57BL/6
gender: female
age: six to eight weeks
tissue: whole blood
treatment: 1Gy gamma rays
time point: 16hr
|
| Treatment protocol |
Whole body irradiation done with X-rays using the Small Animal Radiation Research Platform (SARRP Xstrahl Ltd.). Mice were placed in plastic containers and exposed to a single surface dose of 0-12 Gy with a dose rate of 1.05 Gy/min. Control mice (0 Gy) were placed in the same plastic container as the irradiated mice and sham irradiated. 3 animals/dose/time-point have been included in the study.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted and purified using Ribopure (Ambion) RNA isolation.
|
| Label |
Alexa 555
|
| Label protocol |
Labeled cRNA was prepared from total RNA samples. Briefly, the Poly(A)+ RNA population within total RNA was amplified using MessageAMP II (Applied Biosystems, Foster City, CA). After a second round of reverse transcription, second-strand cDNA synthesis, and purification of double-stranded cDNA, in vitro transcription was performed using T7 RNA polymerase in the presence of Biotin-11-UTP. The quantity and quality of the cRNA were assayed by spectrophotometry and on the Agilent Bioanalyzer.
|
| |
|
| Hybridization protocol |
1 µg of purified cRNA was fragmented to uniform size and applied to Agilent Whole Mouse Genome microarrays 8x60K v1 Design ID 028005 or v2 Design ID 074809 (Agilent Technologies, Santa Clara, CA) in hybridization buffer. Arrays were hybridized at 65° C for 17 hrs. in a rotating incubator, washed at 37° C for 1 min, and stained with Streptavidin-Alexa555.
|
| Scan protocol |
Rinsed and dried arrays were scanned with an Agilent G2505C Microarray Scanner (Agilent Technologies, Santa Clara, CA) at 5 µm resolution.
|
| Description |
16hr_1Gy_3
|
| Data processing |
Agilent Feature Extraction software was used to process the scanned images from arrays (gridding and feature intensity extraction) and the data generated for each probe on the array was analyzed with GeneSpring GX v7.3.1 software (Agilent Technologies, Santa Clara, CA). Only the 30,000+ probes that are printed on both arrays (Agilent-028005, Agilent-074809) are reported.
|
| |
|
| Submission date |
Sep 21, 2017 |
| Last update date |
May 31, 2018 |
| Contact name |
Michael Falduto |
| E-mail(s) |
mfalduto@genusbiosystems.com
|
| Phone |
847-291-9602
|
| Organization name |
GenUs BioSystems, Inc.
|
| Street address |
1808 Janke, Unit M
|
| City |
Northbrook |
| State/province |
IL |
| ZIP/Postal code |
60062 |
| Country |
USA |
| |
|
| Platform ID |
GPL21163 |
| Series (1) |
| GSE104121 |
Radiation-induced molecular alterations after whole body irradiation in a mouse model |
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