|
| Status |
Public on Jul 07, 2016 |
| Title |
LLC1 10Gy irradiation replicate 1 |
| Sample type |
RNA |
| |
|
| Source name |
LLC1 cells irradiated with 10Gy
|
| Organism |
Mus musculus |
| Characteristics |
treatment: single dose of 10Gy
cell line: LLC1
|
| Growth protocol |
LLC1 cells were cultured in DMEM cell culture medium (Gibco) supplemented with 10% fetal bovine serum (Gibco), 2nM glutamine (Gibco), 100 UI/ml penicillin (ROTH) and 0.1 mg/ml streptomycin (ROTH) at 37°C in a humidified atmosphere containing 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA enrich in small noncoding RNAs was isolated from harvested cells using mirVana miRNA Isolation Kit (Ambion) according to manufacturer’s instructions. The quantity and quality of RNA were evaluated using Nanodrop 2000c (Thermo Scientific) and Bioanalyzer (Agilent).
|
| Label |
Cy3
|
| Label protocol |
miRNA labeling was performed using miRNA Complete Labeling and Hyb Kit (Agilent Technologies) according to manufacturer’s instructuctions. In brief, 100 ng of total RNA were defosforilated and directly labeled with Cy3. Samples were dried out and resuspended in Hi-RPM Hybridization Buffer (Agilent Technologies), containing GE Blocking Agent (Agilent, USA), denaturated by heating 5 min in 100°C.
|
| |
|
| Hybridization protocol |
100 ng of RNA labeled with Cy3 dye was prepared for hybridization using Gene Expression Hybridization Kit following manufacturer's protocol (Agilent technologies). Samples were hybridized to Mouse miRNA Expression 8x15k Oligonucleotide Microarrays (Agilent Technologies) for 20 hours at 55°C in rotating hybridization oven.
|
| Scan protocol |
Microarrays were scanned using SureScan Microarray Scanner (Agilent Technologies)
|
| Description |
Biological replicate 1 of 3. Total RNA isolated from LLC1 cells irradiated with single 10 Gy X-ray dose.
|
| Data processing |
Image analysis was performed using Feature Extraction v10.7.3.1 (Agilent Technologies). Raw probe value normalization: Experimental Samples were Scaled to mean of All Samples
|
| |
|
| Submission date |
Jul 06, 2016 |
| Last update date |
Jul 07, 2016 |
| Contact name |
Vaidotas Stankevicius |
| E-mail(s) |
vaidotas.stankevicius@nvi.lt
|
| Phone |
+37068313680
|
| Organization name |
National Cancer Institute
|
| Lab |
Laboratory of molecullar oncology
|
| Street address |
Santariskiu 1
|
| City |
Vilnius |
| State/province |
Lithuania |
| ZIP/Postal code |
LT-08406 |
| Country |
Lithuania |
| |
|
| Platform ID |
GPL7732 |
| Series (2) |
| GSE84109 |
miRNA expression profile of of mouse Lewis lung carcinoma cell line treated with single or fractionated ionizing radiation doses |
| GSE84111 |
Mouse Lewis lung carcinoma cell line treated with single or fractionated ionizing radiation doses nated ionizing radiation doses |
|
