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Status |
Public on Mar 03, 2017 |
Title |
MCF-7_BPA_rep3 |
Sample type |
RNA |
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Source name |
Breast cancer cell line
|
Organism |
Homo sapiens |
Characteristics |
cell line: MCF-7 exposure time: 48h experimental group: Bisphenol A treatment: Bisphenol A concentration (nm): 400
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Treatment protocol |
MCF-7 cells were seeded into 96-well plates with DMEM medium containing 10% FBS at a density of 20,000 cells per well. After 24 h of steroid deprivation in hormone free medium (charcoal stripped FBS), the cells were stimulated with test substances for 48 h in triplicate in three independent experiments.
|
Extracted molecule |
total RNA |
Extraction protocol |
RNA extraction was performed using the Agencourt RNAdvance Cell V2 kit according to the manufacturer's instructions.
|
Label |
Biotin
|
Label protocol |
Single Primer Isothermal Amplified (SPIA) cDNA was generated using the Ovation Pico WTA System V2 kit from Nugen, following the manufacturer's instructions. The SPIA cDNA was fragmented and Biotin-labelled using the Encore Biotin Module from Nugen according to the manufacturer instructions.
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Hybridization protocol |
Hybridisation cocktails were prepared of the fragmented labelled-cDNA according to Nugen's recommendations and hybridised at 45°C overnight. The arrays were washed and stained using wash protocol FS450_0001 recommended for Affymetrix Human Transcriptome 2.0 Arrays on the GeneChip Fluidics station 450.
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Scan protocol |
Arrays were scanned using the GCS3000 7G scanner (Affymetrix) following manufacturer's protocols.
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Data processing |
CEL files were imported into Expression Console software (Affymetrix) and normalised using RMA-sketch (gene-level).
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Submission date |
Aug 09, 2016 |
Last update date |
Mar 06, 2017 |
Contact name |
Robin Mesnage |
E-mail(s) |
robinmes@gmail.com
|
Phone |
+447482206767
|
Organization name |
Kings College London
|
Street address |
Guy's Hospital, 8th Floor, Tower Wing
|
City |
London |
ZIP/Postal code |
SE1 9RT |
Country |
United Kingdom |
|
|
Platform ID |
GPL17586 |
Series (1) |
GSE85350 |
Bisphenol A alternatives can effectively substitute for estradiol in promoting cell growth through estrogen receptor alpha in human breast cancer cells |
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