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Sample GSM665985 Query DataSets for GSM665985
Status Public on Feb 08, 2011
Title Astrocyte_control-transfected_rep1
Sample type RNA
 
Source name mus musculus fetal astrocyte, control-transfected
Organism Mus musculus
Characteristics strain: C57BL/6
age: embryonic day 18
cell type: astrocyte, primary culture
Biomaterial provider Kinki University, Japan
Treatment protocol Astrocytes were transfected in six-well plates (AGC Techno Glass, Japan) containing the medium supplemented with 80 nM each of miR-29 (Ambion Pre-miR hsa-miR-29a/b/c Precursors) or control miRNA (Ambion Pre-miR Negative Control1) using RNAiMax (Invitrogen) in accordance with manufacturer’s protocols. miR-29 RNA for the transfection was prepared by mixing the three miR-29 isoforms in a 1:2:1 molar ratio of Pre-miR-29a:-29b:-29c Precursors. The transfection was conducted three weeks after the initial plating. 48 hr posttransfection, cells were harvested and processed for preparation of RNA samples. The transfection efficiency was above 90% as determined using a fluorochrome-conjugated miR-29 precursor (TAKARA BIO, Japan). RNA extraction was performed in triplicate for each cell type. Total RNA was isolated from the transfected astrocyte cells using an RNAiso Plus (TAKARA BIO). The quality of RNA was assessed using an Agilent 2100 Bioanalyzer, which is indicative of integrity of every RNA sample sufficient for downstream assays.
Growth protocol Primary astrocyte cultures prepared from cerebral cortex of fetal C57BL/6 mouse (embryonic 18-day old) were purchased from ScienCell Research Laboratories. Cells were always cultured and maintained in poly-L-lysine-coated plastic wares (AGC) supplied with Astrocyte Medium (ScienCell) following the supplyer’s recommendations.
Extracted molecule total RNA
Extraction protocol Total RNA was isolated from the transfected astrocyte cells using an RNAiso Plus (TAKARA BIO).
Label Cy3
Label protocol The labeling reaction was carried out separately for the RNA samples using a Quick Amp Labeling Kit one-color, in the presence of cyanine 3-CTP.
 
Hybridization protocol The dye-labeled target (1.6 μg as cRNA) was fragmented and hybridized on an Agilent Whole Mouse Genome 4X44K microarray at 65°C for 17 hr with a Gene Expression Hybridization Kit. The hybridized slide was washed in Gene Expression Wash Buffer 1 at room temperature for 1 min, which was followed by a wash for 1 min in Gene Expression Wash Buffer 2.
Scan protocol The processed microarrays were scanned using an Agilent DNA Microarray Scanner.
Data processing Data extraction from raw image files was done with Agilent Feature Extraction software ver.10.7.
 
Submission date Feb 02, 2011
Last update date Feb 08, 2011
Contact name Hiroyuki Tanabe
Organization name Kinki University
Department Faculty of Agriculture Department of Advanced Bioscience
Street address 3327-204 Nakamachi
City Nara City
State/province Nara
ZIP/Postal code 631-8505
Country Japan
 
Platform ID GPL4134
Series (1)
GSE27035 Global profiling of gene expression in mouse astrocyte in response to the potential longevity determinant miR-29

Data table header descriptions
ID_REF
VALUE Normalized Signal
gSigEval Signal Evaluation, 2:Detected, 1:Un Clear, 0:Not Detected

Data table
ID_REF VALUE gSigEval
12 10 0
13 80.5 2
14 12.4 1
15 78.5 2
16 78.9 2
18 10.1 0
19 27.7 1
20 63.8 2
21 42.5 1
22 2181.4 2
23 14.2 1
24 2351.8 2
25 45 1
26 14 1
27 111.6 2
28 4101.2 2
29 161.5 2
30 10 0
31 9.9 0
32 9.9 0

Total number of rows: 43379

Table truncated, full table size 562 Kbytes.




Supplementary file Size Download File type/resource
GSM665985.txt.gz 2.2 Mb (ftp)(http) TXT
Processed data included within Sample table

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