|
Status |
Public on Oct 01, 2015 |
Title |
MCF-10Acells_untreated_2_cy3 |
Sample type |
genomic |
|
|
Channel 1 |
Source name |
Mammary epithelium
|
Organism |
Homo sapiens |
Characteristics |
cell line: MCF-10A
|
Treatment protocol |
Cells were untreated.
|
Growth protocol |
MCF-10A cells were cultured in a 1:1 mix of DMEM:F-12 media supplemented with 5% horse serum, 1% pen/strep, 10 ug/ml insulin, 0.5 ug/ml hydrocortisone, 20 ng/ml EGF, and 100 ng/ml cholera toxin. Cells were incubated at 37oC, and 5% CO2.
|
Extracted molecule |
genomic DNA |
Extraction protocol |
DNA was purified using a Qiagen Gentra Puregene Cell kit.
|
Label |
cy3
|
Label protocol |
Dye swap labelling of 2ug of test and reference DNA was preformed according to directions supplied by Perkin Elmer for the Constitutional Chip 4.0.
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|
|
Channel 2 |
Source name |
Reference
|
Organism |
Homo sapiens |
Characteristics |
reference: Pooled reference DNA
|
Treatment protocol |
Cells were untreated.
|
Growth protocol |
MCF-10A cells were cultured in a 1:1 mix of DMEM:F-12 media supplemented with 5% horse serum, 1% pen/strep, 10 ug/ml insulin, 0.5 ug/ml hydrocortisone, 20 ng/ml EGF, and 100 ng/ml cholera toxin. Cells were incubated at 37oC, and 5% CO2.
|
Extracted molecule |
genomic DNA |
Extraction protocol |
DNA was purified using a Qiagen Gentra Puregene Cell kit.
|
Label |
cy5
|
Label protocol |
Dye swap labelling of 2ug of test and reference DNA was preformed according to directions supplied by Perkin Elmer for the Constitutional Chip 4.0.
|
|
|
|
Hybridization protocol |
Hybridization was according to the Perkin Elmer protocol. Slides were incubated for 16 hours in Corning
|
Scan protocol |
The slides were scanned on a GenePix 4200A microarray scanner at a resolution of 5 um.
|
Data processing |
Data were analyzed using oneClickCGH software. The following analysis parameters were selected: Background correction: local; Normalization: LOWESS, pin-wise, X&Y adjusted, var stab, Detection algorithm: CBS v2, P-value: 0.01, Gain:0.25, Loss: -0.25, Minimum clones: 1.
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|
|
Submission date |
Jun 23, 2015 |
Last update date |
Oct 01, 2015 |
Contact name |
Caitlin Mills |
Organization name |
McMaster University
|
Street address |
1280 Main St West
|
City |
Hamilton |
ZIP/Postal code |
L8S 4L8 |
Country |
Canada |
|
|
Platform ID |
GPL20610 |
Series (1) |
GSE70206 |
Genetic characterization of the MCF-10A cell line |
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