|
| Status |
Public on Mar 10, 2015 |
| Title |
LM8_91_2 |
| Sample type |
RNA |
| |
|
| Source name |
Osteosarcoma commercial human cell lines
|
| Organism |
Mus musculus |
| Characteristics |
cell line: LM8
properties: Osteoblastic
replicates: Passage number 91
|
| Treatment protocol |
Dunn, LM8 cells were cultured in in Dulbecco’s Modified Eagle Medium (DMEM containing 4.5 g/l glucose)/Ham F12 (Invitrogen, Carlsbad, CA, USA) (1:1) supplemented with 10% fetal calf serum (FCS), 1 unit/ml penicillin G, and 1 μg/ml streptomycin. The cultures were incubated at 37 ºC in a humidified incubator with 5% CO2
|
| Growth protocol |
Dunn, LM8 cells were cultured at same conditions and at 75% confulence the RNAs were isolated
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using Rneasy MiniKit from Qiagen ( Valencia, CA, USA) following manufacturers instruction. RNA was quantified using a NanoDrop-1000 spectrophotometer and quality was monitored with the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA).
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 100 ng total RNA using the low input quick amplification labeling kit (Agilent) according to the manufacturer's instructions, followed by RNAeasy column purification (QIAGEN, Valencia, CA). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
|
| |
|
| Hybridization protocol |
Mouse GE 4x44K v2 Microarrays were incubated for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent), then immediately air dried prior scanning.
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray C reader using using a scan Control software version A.8.4.1 and one color scan setting for 4x44k array slides (Scan Area 61x21.6 mm, Scan resolution 5 um, Dye channel is set to Green and Green PMT is set to 100%).
|
| Description |
Mouse osteosarcoma cell line
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 10.7.3 (Agilent) using default parameters to obtain background subtracted and spatially detrended Processed Signal intensities.
|
| |
|
| Submission date |
Mar 09, 2015 |
| Last update date |
Mar 10, 2015 |
| Contact name |
Ram Mohan Ram Kumar |
| E-mail(s) |
rkumar@research.balgrist.ch
|
| Organization name |
University Hospital Balgrist, University of Zurich, Zurich, Switzerland
|
| Department |
Orthopaedics
|
| Street address |
Forchstrasse 340
|
| City |
Zurich |
| ZIP/Postal code |
8008 |
| Country |
Switzerland |
| |
|
| Platform ID |
GPL10333 |
| Series (2) |
| GSE66674 |
Identification of genes regulated during passaging of mouse osteosarcoma cell lines |
| GSE66675 |
Identification of genes regulated during passaging of human and mouse osteosarcoma cell lines |
|
