|
| Status |
Public on Feb 01, 2011 |
| Title |
BV2 1 |
| Sample type |
RNA |
| |
|
| Source name |
Immortalized cell line BV2
|
| Organism |
Mus musculus |
| Characteristics |
cell line: BV2
cell type: normal
|
| Treatment protocol |
No treatments were performed. mRNA levels under basal conditions were measured.
|
| Growth protocol |
Cells grown in DMEM with 10% FBS and penicillin/streptomycin at 37C with 5% CO2 humified air.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from cells using TRIzol reagent (Invitrogen) under RNase-free conditions, followed by chloroform treatment (100ul/ml). After centrifugation, the aqueous phase containing total RNA is then extracted and treated with isopropanol (500uL/mL) for 5-10 mins. After centrifugation, the RNA pellet was isolated, washed with ethanol (75%; 1ml/1mL of reagent solution), followed by another centrifugation and air-dried. The RNA pellet was resuspended in RNase-free water. Total RNA quantity was assessed using NanoDrop ND-1000, and the Bioanalyzer 2100 determined the 28S/18S ratio for quality assessment.
|
| Label |
Cy3
|
| Label protocol |
To synthesize targets for hybridization, we began with 10.0 µg of total RNA and used Invitrogen's SuperScript Double-Stranded cDNA Synthesis kit using oligo (dT) primer (Promega) followed by DNA labeling using 1 O.D. Cy3-labeled random nonamer primer (TriLink Biotechnologies) and 100U Klenow fragment 3’>5’ exo- (NEB) per 1.0 µg double-stranded cDNA.
|
| |
|
| Hybridization protocol |
Single-color hybridization, post-hybridization washing and scanning were done with NimbleGen reagents and according to the NimbleGen's User's Guide for Expression Analysis.
|
| Scan protocol |
Images were acquired using a GenePix 4200A scanner with GenePix 6.0 software.
|
| Description |
Biological replicate 1 of 4.
|
| Data processing |
The data from these arrays were extracted using the software NimbleScan 2.4 (Roche NimbleGen) and processed into PAIR files. The data was not normalized.
|
| |
|
| Submission date |
Dec 06, 2010 |
| Last update date |
Jun 25, 2012 |
| Contact name |
Jacqueline Lopez |
| E-mail(s) |
jacqueline.ann.lopez@gmail.com
|
| Phone |
574-631-1902
|
| Organization name |
University of Notre Dame
|
| Department |
Biology
|
| Lab |
Pfrender Laboratory
|
| Street address |
019 Galvin Life Science Center
|
| City |
Notre Dame |
| State/province |
Indiana |
| ZIP/Postal code |
46556 |
| Country |
USA |
| |
|
| Platform ID |
GPL11059 |
| Series (2) |
| GSE25886 |
Analysis of mRNA levels of GPCR and GPCR-related signaling proteins in 3 mouse cell lines: AtT20, BV2 and N18 |
| GSE25901 |
Expression of G protein-coupled receptors and related proteins in HEK293, AtT20, BV2, and N18 cell lines as revealed by microarray analysis |
|
