|
| Status |
Public on Apr 10, 2019 |
| Title |
Allogeneic transplant - MC-deficient mouse 3 |
| Sample type |
RNA |
| |
|
| Source name |
skin after bone marrow transplant
|
| Organism |
Mus musculus |
| Characteristics |
transplant type: Allogeneic
strain: B6.Cg-KitW-sh/HNihrJaeBsmGlliJ
|
| Treatment protocol |
Recipient mice were irradiated with 850cGy ionizing radiation, then injected IV with 5e6 LP/J bone marrow cells and 2e6 LP/J splenocytes. GVHD symptoms were allowed to progress for 7 weeks, then mice were euthanized by CO2 and skin was isolated from the nape of the neck and stored in RNAlater solution at -20C
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Skin in RNAlater solution was transferred to 1ml of Trizol, then minced and homogenized. Trizol extraction of RNA was performed as per the manufacturer's instructions. RNA was assessed for quality by Agilent Bioanalyzer, then measured by QuBit and 100ng was loaded into the Myleoid Innate Immunity kit.
|
| Label |
N/A
|
| Label protocol |
RNA was assessed for quality by Agilent Bioanalyzer, then measured by QuBit and 100ng was loaded into the NanoString Myleoid Innate Immunity kit.
|
| |
|
| Hybridization protocol |
In the NanoString kit, individual mRNA molecules are directly hybridized with barcoded fluorescent molecules per the manufacturer's instructions.
|
| Scan protocol |
Samples were scanned by a Nanostring Sprint system and number of barcoded spots was counted directly.
|
| Description |
Total RNA was extracted from skin taken from the nape of the neck. RNA was assessed for quality by Bioanalyzer and 100ng was loaded into the Nanostring Myeloid Innate Immunity Panel.
|
| Data processing |
Data was assessed using the NanoString nSolver software, which normalizes the readouts to a panel of ~20 housekeeping genes.
|
| |
|
| Submission date |
Mar 22, 2019 |
| Last update date |
Apr 11, 2019 |
| Contact name |
Ethan Jesse Strattan |
| E-mail(s) |
ethan.strattan@uky.edu
|
| Organization name |
University of Kentucky
|
| Department |
Hematology/BMT
|
| Lab |
Hildebrandt
|
| Street address |
900 South Limestone Street, Wethington Building, CTW453
|
| City |
Lexington |
| State/province |
KY |
| ZIP/Postal code |
40508 |
| Country |
USA |
| |
|
| Platform ID |
GPL25266 |
| Series (1) |
| GSE128704 |
Analysis of skin in wild-type vs. mast cell-deficient mice after allogeneic bone marrow transplant |
|
