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Status |
Public on Jun 01, 2010 |
Title |
FHCE1 vs IHCE1 human epididymal cells replicate 2 |
Sample type |
RNA |
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|
Channel 1 |
Source name |
FHCE1
|
Organism |
Homo sapiens |
Characteristics |
cell line: FHCE1 tissue: epididymis cell type: SV40 immortalized primary cell line fertility status: fertile
|
Growth protocol |
Cells were grown to confluence on inserts (Corning) in a humified chamber (32°C, 5% CO2).
|
Extracted molecule |
total RNA |
Extraction protocol |
Total cellular RNA was isolated using the Illustra RNAspin Mini kit (GE Healthcare, Buckinghamshire, Germany) according to the manufacturer’s instructions. The quality of the total RNA was verified using the Agilent 2100 Bioanalyzer (Agilent Technologies).
|
Label |
Cy3
|
Label protocol |
Fluorescent cRNA were prepared from 500ng of total RNA using the Agilent Low RNA Input Linear Amplification Kit. As a last step of the extraction procedure, the cRNA was purified with the Rneasy Mini-Kit (Qiagen).
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Channel 2 |
Source name |
IHCE1
|
Organism |
Homo sapiens |
Characteristics |
cell line: IHCE1 tissue: epididymis cell type: SV40 immortalized primary cell line fertility status: obstructive azoospermic patient
|
Growth protocol |
Cells were grown to confluence on inserts (Corning) in a humified chamber (32°C, 5% CO2).
|
Extracted molecule |
total RNA |
Extraction protocol |
Total cellular RNA was isolated using the Illustra RNAspin Mini kit (GE Healthcare, Buckinghamshire, Germany) according to the manufacturer’s instructions. The quality of the total RNA was verified using the Agilent 2100 Bioanalyzer (Agilent Technologies).
|
Label |
Cy5
|
Label protocol |
Fluorescent cRNA were prepared from 500ng of total RNA using the Agilent Low RNA Input Linear Amplification Kit. As a last step of the extraction procedure, the cRNA was purified with the Rneasy Mini-Kit (Qiagen).
|
|
|
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Hybridization protocol |
0.75 mg of cRNA were hybridized to the Agilent microarrays (Agilent Human 1A Microarray (V2) G411B) using the Agilent In Situ Hybridization Kit Plus) according to the protocol provided by the manufacturer.
|
Scan protocol |
Following hybridization, microarrays were scanned with a ScanArray Express scanner (Perkin Elmer). Fluorescence ratios for array elements were extracted using the ScanArray Express Software (Perkin Elmer) and imported into the GeneSpring 6.1 software (Agilent Technologies) for further analysis.
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Description |
Replicate 2 of 3, IHCE1 vs FHCE1, dye swap
|
Data processing |
GeneSpring Software 6.1 (Agilent) was used for analysis following LOWESS normalization. A dye swap was applied to some of the samples (CT1neg-CL1neg and CT2neg-CL2neg).
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Submission date |
Apr 19, 2010 |
Last update date |
Apr 19, 2010 |
Contact name |
Daniel Cyr |
E-mail(s) |
daniel.cyr@iaf.inrs.ca
|
Phone |
514-630-8833
|
Fax |
514-630-8850
|
Organization name |
INRS-Institut Armand-Frappier
|
Street address |
245, Boulevard Hymus
|
City |
Pointe-Claire |
State/province |
Québec |
ZIP/Postal code |
H9R1G6 |
Country |
Canada |
|
|
Platform ID |
GPL6087 |
Series (1) |
GSE21391 |
Comparison of gene expression between a human epididymal cell line derived from the caput epididymidis of a fertile patient and another one derived from the caput epididymidis of an obstructive azoospermic patient |
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