|
| Status |
Public on May 25, 2011 |
| Title |
ZR75, technical replicate 1 |
| Sample type |
RNA |
| |
|
| Source name |
ZR75 is a breast cancer cell line
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: ZR75
cell type: breast cancer
genotype: ER positive ZR-75-1/GFP/puro
|
| Growth protocol |
MDA-MB-435-F-L/GFP cells, which are highly invasive and metastatic variants of human carcinoma MDA-MB-435 cells, were isolated in our laboratory. The human breast cancer cell line ZR-75-1 was originally obtained from the American Type Culture Collection (Manassas, VA) and stably transfected with the enhanced green fluorescent protein (GFP) for the detection of metastases in vivo with green fluorescence imaging. We used a clone of GFP-transfected MDA-MB-231 cells named clone 10 from Dr. Julie Anne Sterling at Vanderbilt University, which was resistant to G418 antibiotic.
Cell lines were cultured in McCoy's 5A medium supplemented with pyruvate, vitamins, amino acids, antibiotics, and 10% fetal bovine serum. Working cultures were maintained at 37°C in a humidified incubator with 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol (Invitrogen) and RNeasy (Qiagen) kits were used to extract total RNA according to the manufacturer's instructions.
|
| Label |
Biotin
|
| Label protocol |
Per RNA labeling, 5 micrograms of total RNA was used in conjunction with the Affymetrix recommended protocol One-Cycle Target Labeling and Control Reagents (Catalog# 900493 ).
|
| |
|
| Hybridization protocol |
The hybridization cocktail containing the fragmented and Biotin labeled cDNAs was hybridized to the Affymetrix Mouse 430 2.0 GeneChip. The chips were washed and stained by the Affymetrix Fluidics Station using the standard format and protocols as described by Affymetrix. The probe arrays were stained with streptavidin phycoerythrin solution (Molecular Probes, Carlsbad, CA) and enhanced by using an antibody solution containing 0.5 mg/mL of biotinylated anti-streptavidin (Vector Laboratories, Burlingame, CA).
|
| Scan protocol |
Genechips were scanned using an Affymetrix Gene Chip Scanner 3000.
|
| Description |
LZ07_ZR75_1
|
| Data processing |
Gene expression intensities were calculated using GeneChip® Command Console® Software (AGCC) and Expression Console™ Software. RMA normalization was performed to generate the CHP files. Genesifter software and Pairwise comparisons were perfomed with Benjamini and Hochberg correction.
|
| |
|
| Submission date |
Feb 24, 2011 |
| Last update date |
May 25, 2011 |
| Contact name |
abdel G Elkahloun |
| E-mail(s) |
[email protected]
|
| Phone |
301 402 3170
|
| Organization name |
NHGRI-NIH
|
| Lab |
MICROARRAY CORE
|
| Street address |
50, SOUTH DRIVE
|
| City |
BETHESDA |
| ZIP/Postal code |
20892 |
| Country |
USA |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE27515 |
Isolation and characterization of a metastatic hybrid cell line generated by ER negative and ER positive breast cancer cells in mouse bone marrow |
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