|
| Status |
Public on Jul 16, 2019 |
| Title |
A375 Parental, biological rep2 |
| Sample type |
RNA |
| |
|
| Source name |
melanoma cell line derived from Lymph Node Metastasis
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: A375
sample type: BRAFV600E parental cells sensitive to BRAF inhibitors
phenotype: Parental cell line
|
| Treatment protocol |
A375 and A375R cells were removed from culture, seeded into 10-cm dishes, and maintained in CM for 48 h. The cells were then detached with a solution of EDTA in PBS, washed in PBS and frozen in a RNA-later®/PBS solution.
|
| Growth protocol |
A375 cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum, 2 mM L-glutamine and 50 m/ml gentamicin (Complete Medium, CM). The A375R cell line was maintained in CM supplemented with 1.5 mM dabrafenib
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using TRIzol reagent (Invitrogen) and mirVana miRNA isolation kit (Ambion) following the manufacturers protocol. RNA quantity and quality were determined using a Nanodrop-1000 spectrophotometer (Nanodrop Technology) and Agilent 2100 Bioanalyzer.
|
| Label |
biotin
|
| Label protocol |
130-1000 ng of total of RNA was biotin labeled using the FlashTag™ HSR RNA labeling kit for Affymetrix GeneChip™ miRNA 3.1 Array Strip hybidization.
|
| |
|
| Hybridization protocol |
Following biotinylation, these fragments were hybridized for 20 hr at 45°C on Affymetrix miRNA 3.1 array strips in GeneAtlas Hybridization Station. GeneChips were washed and stained using GeneAtlas Hybridization, Wash, and Stain Kit for miRNA Array strips.
|
| Scan protocol |
GeneChip were finally scanned using GeneAtlas Scanner.
|
| Data processing |
Affymetrix Expression Console. Data were preprocessed following the RMA procedure using quantile normalization
|
| |
|
| Submission date |
Jul 25, 2018 |
| Last update date |
Jul 16, 2019 |
| Contact name |
Adriana Agnese Amaro |
| E-mail(s) |
adriana.amaro@hsanmartino.it
|
| Phone |
+390105558312
|
| Organization name |
IRCCS Ospedale Policlinico SAN MARTINO - IST - NATIONAL CANCER RESEARCH INSTITUTE
|
| Department |
Molecular Pathology
|
| Street address |
Largo Rosanna Benzi 10
|
| City |
Genoa |
| ZIP/Postal code |
16132 |
| Country |
Italy |
| |
|
| Platform ID |
GPL16384 |
| Series (1) |
| GSE117666 |
miRNA expression profiling of the A375 melanoma cell line and its dabrafenib-resistant A375R sub-line. |
|
