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Status |
Public on Feb 25, 2009 |
Title |
Cortex 15WT |
Sample type |
RNA |
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Source name |
cerebral cortex, wild-type
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Organism |
Mus musculus |
Characteristics |
genotype: Cux2+/- tissue: brain gender: male
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Extracted molecule |
total RNA |
Extraction protocol |
Cerebral cortex were dissected and total RNA was obtained using Qiagen RNAeasy kit.
|
Label |
Cy3
|
Label protocol |
For each sample, we have generated cyanine 3-labeled cRNA from one microgram of total RNA, using the Agilent Low RNA Input Linear Amplification Kit PLUS, One-Color. Briefly, total RNA is reverse-transcribed using T7-oligo (dT) primer; after first and second strand cDNA synthesis, cRNA is synthesized from the double-stranded cDNA using T7 RNA polymerase, which simultaneously incorporates cyanine 3.
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Hybridization protocol |
1.65 ug of Cy3 labelled aRNA were hybridized for 17 hours at 65ºC in a hybridization oven (G2545A, Agilent) set to 10 rpm in a final concentration of 1X GEx Hybridization Buffer HI-RPM, according to manufacturer's instructions (One-Color Microarray-Based Gene Expression Analysis, Agilent Technologies).
|
Scan protocol |
Arrays were scanned at 5um resolution on an Agilent DNA Microarray Scanner (G2565BA, Agilent Technologies) using the default settings for 4x44k format one-color arrays. Images provided by the scanner were analyzed using Feature Extraction software (Agilent Technologies).
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Description |
WT
|
Data processing |
Data files from Feature Extraction were imported into GeneSpring® GX software version 9.0. (Agilent Technologies). Quantile normalization was performed and expression values (log2 transformed) were obtained for each probe.
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|
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Submission date |
Feb 24, 2009 |
Last update date |
Feb 24, 2009 |
Contact name |
Ana Dopazo |
E-mail(s) |
adopazo@cnic.es
|
Phone |
34914531217
|
Organization name |
CNIC
|
Department |
Genomics Unit
|
Street address |
Melchor Fdez Almagro
|
City |
madrid |
ZIP/Postal code |
28029 |
Country |
Spain |
|
|
Platform ID |
GPL7202 |
Series (1) |
GSE14971 |
Comparative gene expression profile of Cux2 knock-out cortex |
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