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Sample GSM725732 Query DataSets for GSM725732
Status Public on Jul 04, 2011
Title B9 antimir-neg
Sample type RNA
Source name hMADS cells, B9, antimir-neg
Organism Homo sapiens
Characteristics cell type: human adipose tissue-derived stem cells
clone: B9
agent: control antimiR
Treatment protocol All transfections were performed with HighPerfect transfection reagent (Qiagen). Adipogenic medium was composed of DMEM/Ham’s F12 media supplemented with 10 µg/ml transferrin, 0.86 µM insulin, 0.2 nM triiodothyronine, 1 µM dexamethasone, 100 µM isobutyl-methylxanthine and 1 µM rosiglitazone. Three days later, the medium was changed (dexamethasone and isobutyl-methylxanthine were omitted).
Growth protocol Cells were maintained in the proliferation medium, which is composed of DMEM (low glucose) containing 10% fetal calf serum (FCS), 0.01 M HEPES, 100 U/ml penicillin and streptomycin.
Extracted molecule total RNA
Extraction protocol Total RNA was extracted with the Qiagen Rneasy kit. RNA was quantified using a NanoDrop-1000 spectrophotometer and quality was monitored with the Agilent 2100 Bioanalyzer (Agilent).
Label Cy3
Label protocol Cyanine-3 (Cy3) labeled cRNA was prepared from 200 ng RNA using the Low Input Quick Amp Labeling Kit, one-color (Agilent) according to the manufacturer's instructions, followed by RNAeasy column purification (QIAGEN, Valencia, CA). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
Hybridization protocol 600 ng of Cy3-labelled cRNA (specific activity >10.0 pmol Cy3/µg cRNA) was fragmented at 60°C for 30 minutes in a reaction volume of 250 ml containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturer's instructions. On completion of the fragmentation reaction, 250 ml of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent SurePrint G3 Human GE 8x60K for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent).
Scan protocol Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2565) using one color scan setting for 8x60k array slides (Scan Area 61x21.6 mm, Scan resolution 3µm, Dye channel is set to Green and Green PMT is set to 100%).
Description Gene expression after 4 days in adipogenic medium.
Data processing The scanned images were analyzed with Feature Extraction Software (Agilent) using default parameters (protocol GE1_107_Sep09 and Grid: 028004_D_20101102) to obtain background subtracted and spatially detrended Processed Signal intensities.
Submission date May 17, 2011
Last update date Jul 04, 2011
Contact name Kevin Lebrigand
Organization name IPMC/CNRS
Lab Functional Genomics Platform of Nice-Sophia-Antipolis, France.
Street address 660 route des lucioles
City Valbonne - Sophia-Antipolis
ZIP/Postal code 06560
Country France
Platform ID GPL13607
Series (1)
GSE29207 Role of miR-30 miRNAs during adipogenesis

Data table header descriptions
VALUE Normalized signal intensity

Data table
1 16.67834636
2 5.100136671
3 5.236492618
4 7.289096702
5 8.769755213
6 5.966937979
7 11.50908219
8 11.53035756
9 5.43629512
10 6.370687407
11 5.352264173
12 9.304693557
13 8.895120819
14 9.310385568
15 10.86395393
16 5.059614856
17 6.4749227
18 5.135452784
19 5.785288981
20 9.964882318

Total number of rows: 62976

Table truncated, full table size 1086 Kbytes.

Supplementary file Size Download File type/resource
GSM725732.txt.gz 12.1 Mb (ftp)(http) TXT
Processed data included within Sample table

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