|
| Status |
Public on Apr 08, 2014 |
| Title |
C19.6 |
| Sample type |
RNA |
| |
|
| Source name |
HTR-8/SVeo cell line, transfected with modified BAC, clone # 6
|
| Organism |
Homo sapiens |
| Characteristics |
treatment: transfected with modified BAC, clone # 6
cell type: HTR-8/SVneo
tissue: Placenta
|
| Treatment protocol |
HTR-8/SVeo were transfected with a BAC (RP11-1055O17) that was modified by recombineering in order to insert a GFP-zeocin cassette. After transfection using PEI (Polyethylenimine), selection was carried out for two weeks with 200 μg/ml Zeocin. Resistant clones were selected and expanded.
|
| Growth protocol |
HTR-8/SVeo cells were cultured in DMEM with 5% BGS (Bovine Growth Serum, Hyclone) and 1% Pen/Strep. Primary trophoblasts were isolated from third trimester placentas after Trypsin/Dispase/DNaseI digestion followed by purification on a Percoll gradient. Primary trophoblasts were cultured for 48h to 72h in DMEM supplemented with 10% FBS (Fetal Bovine Serum), 20 mM Hepes ph 7.4, and a mixture of penicilin-streptomycin-fungizone. Cells were maintained at 37ºC/5% CO2
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA were prepared using miRNeasy Mini Kit following the manufacturer's instructions
|
| Label |
Cy3
|
| Label protocol |
The labeling was performed using Low Input Quick Amp Labeling Kit, One-Color from Agilent following the manufacturer's instructions
|
| |
|
| Hybridization protocol |
The hybridization was performed using Gene Expression Hybridization Kit and Gene Expression Wash Buffer Kit from Agilent following the manufacturer's instructions
|
| Scan protocol |
Slides were scanned on the Agilent G4900DA SureScan Microarray Scanner System using the protocol GE1_1100_Jul11 (Scan Area 61x21.6 mm, Scan resolution 5 um, Dye channel is set to Green and Green PMI gain is set to100%)
|
| Description |
Gene expression profiles in C19MC transfected HTR-8/SVeo cells
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 11.0.1.1 (Agilent) using default parameters (protocol GE1_1100_Jul11 and Grid: 028004_D_F_20120411)
|
| |
|
| Submission date |
Apr 07, 2014 |
| Last update date |
Apr 08, 2014 |
| Contact name |
Jean-Francois Mouillet |
| E-mail(s) |
mouilletj@mwri.magee.edu
|
| Phone |
412-641-1684
|
| Organization name |
University of Pittsburgh
|
| Department |
MWRI - OBGYN and Reproductive Sciences
|
| Lab |
Sadovsky lab
|
| Street address |
204 Craft Ave
|
| City |
Pittsburgh |
| State/province |
PA |
| ZIP/Postal code |
15206 |
| Country |
USA |
| |
|
| Platform ID |
GPL13607 |
| Series (2) |
| GSE56564 |
Induced expression of the C19MC miRNAs in HTR-8/SVneo cells (mRNA) |
| GSE56565 |
Induced expression of the C19MC miRNAs in HTR-8/SVneo cells |
|
