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Status |
Public on Feb 06, 2007 |
Title |
Wild-type (14.5BS) a(16012201010028) |
Sample type |
RNA |
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Channel 1 |
Source name |
Wild-type (14.5BS) a
|
Organism |
Mus musculus |
Characteristics |
Strain: C57BL/6J Gender: male Age: 14.5dpc Development stage: embryo
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Biomaterial provider |
Chang-Yi Cui
|
Growth protocol |
The UMRR is provided in a solution of 70% ethanol and 0.1 M sodium acetate. Prepare the UMRR for use as follows: 1.Centrifuge the tube at 12,000 x g for 15 minutes at 4C. 2.Carefully remove the supernatant. 3.Wash the pellet in 70% ethanol. 4.Centrifuge the tube at 12,000 x g for 15 minutes at 4C. 5.Carefully remove the supernatant and air-dry the pellet at room temperature for 30 minutes to remove retained ethanol. 6.Resuspend the pellet in RNase-free water to the desired concentration.
|
Extracted molecule |
total RNA |
Extraction protocol |
RNA samples from embryonic skin were subjected to LiCl precipitation after Triazol isolation to eliminate genomic DNA contamination. LiCl precipitation was effective for cleaning RNA samples, which is proven in real-time PCR.
|
Label |
Cy3
|
Label protocol |
Total RNA was labeled using Agilents Fluorescent Linear Amplification kit, according to manufacturers instructions (Product Number G2554A or P/N G2556-66002, Version 3.0, June 2002).
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Channel 2 |
Source name |
Universal Mouse Reference RNA
|
Organism |
Mus musculus |
Characteristics |
Universale Mouse Reference
|
Growth protocol |
The UMRR is provided in a solution of 70% ethanol and 0.1 M sodium acetate. Prepare the UMRR for use as follows: 1.Centrifuge the tube at 12,000 x g for 15 minutes at 4C. 2.Carefully remove the supernatant. 3.Wash the pellet in 70% ethanol. 4.Centrifuge the tube at 12,000 x g for 15 minutes at 4C. 5.Carefully remove the supernatant and air-dry the pellet at room temperature for 30 minutes to remove retained ethanol. 6.Resuspend the pellet in RNase-free water to the desired concentration.
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was extracted from cultured cells using a Stratagene Absolutely RNA kit. DNase-treated total RNA samples from 11 cultured cell lines derived from embryo, embryo fibroblast, kidney, liver/hepatocyte, lung/alveolar macrophage, B-lymphocyte, T-lymphocyte (thymus), mammary gland, muscle myoblast, skin, and testis were pooled in equal parts
|
Label |
Cy5
|
Label protocol |
Total RNA was labeled using Agilents Fluorescent Linear Amplification kit, according to manufacturers instructions (Product Number G2554A or P/N G2556-66002, Version 3.0, June 2002).
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Hybridization protocol |
Agilent 60-mer oligo microarray processing protocol (SSC Wash/SureHyb Chamber set-up) V4.1, April 2004. Agilent Publication Number: G4140-90030 V4.1 APRIL 2004. The manual can be found at https://www.chem.agilent.com/scripts/literaturePDF.asp?iWHID=34961, or through a WEB search on the manual title.
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Scan protocol |
Slides were scanned with an Agilent DNA Microarray Scanner model G2505-64120 at 100% PMT in both channels, with a scan resolution of 10um. A scan window of 61 x 21.6 mm was used, then images were cropped to size and saved in a modified two-color TIFF format. Images were examined visually for evidence of foreign debris or major failures. Agilent Microarray Scanner User Manual (6.3) http://www.chem.agilent.com/scripts/literaturePDF.asp?iWHID=33696
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Description |
Wild-type (14.5BS)
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Data processing |
Data are extracted with Agilent Feature Extraction Software.The data were further processed with NIA ANOVA tool utilities.See http://lgsun.grc.nia.nih.gov/ANOVA for details.
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Submission date |
Feb 05, 2007 |
Last update date |
Feb 05, 2007 |
Contact name |
Minoru S.H. Ko |
E-mail(s) |
kom@mail.nih.gov
|
Phone |
410-558-8359
|
Organization name |
NIH
|
Department |
National Institute on Aging
|
Lab |
Lab of Genetics
|
Street address |
251 Bayview Blvd, Suite 100, 10C
|
City |
Baltimore |
State/province |
MD |
ZIP/Postal code |
21224 |
Country |
USA |
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Platform ID |
GPL2550 |
Series (1) |
GSE6952 |
Ectodysplasin regulates the lymphotoxin-beta pathway for hair differentiation |
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