NCBI Logo
GEO Logo
   NCBI > GEO > Accession DisplayHelp Not logged in | LoginHelp
GEO help: Mouse over screen elements for information.
          Go
Sample GSM498998 Query DataSets for GSM498998
Status Public on Jul 05, 2010
Title C2C12_siRNA_Six1_R2
Sample type RNA
 
Source name C2C12, siRNA Six1
Organism Mus musculus
Characteristics cell line: C2C12
cell type: myoblasts
sirna treatment: siSix1
Treatment protocol The cells were transfected, 24h before the start of differentiation, with 100 pmol of siRNA (Dharmacon) using the Lipofectamine2000 (Invitrogen) using the manufacturer's protocol.
Growth protocol Cells were grown in growth medium (GM) DMEM High Glucose (Hyclone); 10% Characterized Fetal Bovine Serum (Hyclone); 1% L-Glutamine (Hyclone); 1% Penicillin / Streptomycin (Hyclone). At the start of differentiation, the cells were grown in differentiation medium (DM) DMEM High Glucose (Hyclone); 2% Horse Serum (Invitrogen); 1% L-Glutamine (Hyclone); 1% Penicillin / Streptomycin (Hyclone).
Extracted molecule total RNA
Extraction protocol Total RNA was isolated with the Absolutely RNA Mini-Prep Kit (Stratagene) following the manufacturer's protocol. RNA quantity and quality was assessed with the NanoDrop ND-1000. Integrity of the RNA was determined by agarose gel.
Label Cy3
Label protocol Cyanine-3 (Cy3) labeled cRNA was prepared from 300ng RNA using the Quick Amp Labeling Kit, one-color kit (Agilent) according to the manufacturer's instructions, followed by RNAeasy column purification (QIAGEN). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
 
Hybridization protocol 1.65 ug of Cy3-labelled cRNA (specific activity >9.0 pmol Cy3/ug cRNA) was fragmented at 60°C for 30 minutes in a reaction volume of 55 ul containing 1x Agilent fragmentation buffer and 25x Agilent blocking agent following the manufacturer's instructions. On completion of the fragmentation reaction, 55 ul of 2x Agilent hybridization buffer was added to the fragmentation mixture and 100 ul of the mixture was hybridized to Agilent Whole Mouse Genome Microarrays (G4122F) 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), then air dried.
Scan protocol Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2565C) using one color scan setting for 1x44k array slides (Scan Area 61x21.6 mm, Scan resolution 5um, 0.10 XDR, Dye channel is set to Green and Green PMT is set to 100%).
Description Gene Expression profiling of C2C12 24h after start of differentiation when cells were transfected with an siRNA sequence against Six1.
Data processing The scanned images were analyzed with Feature Extraction Software 10.5.1.1 (Agilent) using default parameters (protocol GE1-v5_10_Apr08 and Grid: 014868_D_F_20080627) to obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded.

Raw data were processed using GeneSpring GX (Agilent). Using the gMedian signal transformed to log base 2, the probes were normalized within the sample to the 75th percentile and the samples were then normalized by baseline transformation through the median of each sample.
 
Submission date Jan 20, 2010
Last update date May 10, 2023
Contact name Alexandre Blais
E-mail(s) alexandre.blais@uottawa.ca
Organization name University of Ottawa
Street address 451 Smyth Road
City Ottawa
State/province ON
ZIP/Postal code K1H 8M5
Country Canada
 
Platform ID GPL7202
Series (2)
GSE19967 Gene Expression Profiling of siRNA Knock-Down of Six1, Six4 and Myogenin in C2C12
GSE19988 Six1 and Six4 function in myoblast differentiation

Data table header descriptions
ID_REF
VALUE normalized log2 signal

Data table
ID_REF VALUE
GE_BrightCorner -0.8434658
DarkCorner -1.2315049
A_52_P616356 -1.3440619
A_52_P580582 -0.502233
A_52_P403405 -1.1242828
A_52_P819156 -0.6009464
A_51_P331831 -0.88837576
A_51_P430630 -1.2639832
A_52_P502357 -1.1401029
A_52_P299964 -0.4014659
A_51_P356389 -0.939064
A_52_P684402 -0.26576614
A_51_P414208 -1.0664253
A_51_P280918 -0.10617542
A_52_P613688 -1.2356534
A_52_P258194 -1.0508728
A_52_P229271 -1.1792362
A_52_P214630 0.042798042
A_52_P579519 -0.8792255
A_52_P979997 -1.409339

Total number of rows: 41267

Table truncated, full table size 980 Kbytes.




Supplementary file Size Download File type/resource
GSM498998.txt.gz 2.2 Mb (ftp)(http) TXT
Processed data included within Sample table

| NLM | NIH | GEO Help | Disclaimer | Accessibility |
NCBI Home NCBI Search NCBI SiteMap