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Sample GSM1611273 Query DataSets for GSM1611273
Status Public on Apr 21, 2015
Title Postnatal P12, biological rep3
Sample type RNA
Source name mouse trunk muscles including skeletal muscle proliferant satellite cells, P12
Organism Mus musculus
Characteristics strain/background: C57BL/6J
genotype/variation: Pax3GFP/+
tissue: trunk muscles including skeletal muscle proliferant satellite cells
age: postnatal day 12
Treatment protocol Pax3GFP/+ mice of the appropriate age were detected by PCR and splotch white tag in the belly, and GFP confirmed under fluorescent stereoscope. Muscle samples were isolated from the trunk and digested in DMEM high glucose but no phenol red, 0.1% Trypsin, 0.1% Collagenase D and 0.01mg/mL DNaseI, and purified by filtration using 100µm and 40µm cell strainers. GFP cells were stained using propidium iodide to exclude dead cells and purified via FACS Aria II based on gating of GFP signal.
Growth protocol Pax3GFP/+ mice were bred with C57BL/6J and the day where plugs were detected was considered as E0.5 (E, embryonic days).
Extracted molecule total RNA
Extraction protocol RNeasy® Micro Kit (QIAGEN) RNA extraction protocol was used according to the manufacturer's instructions.
Label biotin
Label protocol Biotinylated cRNA were prepared according to the Encore Biotin Module (NuGEN) protocol from 100ng total RNA ( Single-stranded cDNA is fragmented and characterized by chemical and enzymatic methods leading to molecules of about 50 to 100 bases. Next, a biotin-labeled nucleotide is attached to the 3' end of each fragment.
Hybridization protocol High-density oligonucleotide arrays containing 45,000 sets of oligonucleotide probes (25 mers) that cover all 30,000 genes encoded by the murine genome (Affymetrix Mouse Genome 430 2.0 Arrays, Ref 900495) were used for gene expression detection. Hybridization during 16 hours at 45°C in a rotary oven (Affymetrix), washing and staining (GeneChip® Fluidics Station 450) and scanning (GeneChip Scanner 3000) were carried out according to NuGEN and Affymetrix protocols.
Scan protocol GeneChips were scanned using the GeneChip Scanner 3000.
Description Sample name: P-12.C.2006
Data processing Expression Console software (Affymetrix) was used for image analysis and to determine probe signal levels. The quality and statistical analysis of the data were made using the GeneSpring GX11 analysis software (Agilent Technologies).
Data needed to be pre-treated before final analysis to normalize two different sets of arrays (2006 or 2012, as indicated in the sample names).
Expression profiles were normalized in batch using the RMA algorithm (affy R package) yielding a (probe sets, samples) matrix. As the 11 samples were obtained by merging two series, the Combat algorithm (Johnson WE, et al. Biostatistics, 2007) was used to normalize the corresponding batch effect. Expression profiles were aggregated by Gene Symbol (mean across probe sets) using Affymetrix csv annotation file (na32 version).
Submission date Feb 13, 2015
Last update date Apr 21, 2015
Contact name Sonia Alonso-Martin
Phone +33684321566
Organization name Myology Research Center U974-INSERM - UPMC-Paris VI - Institut de Myologie
Department Development and stem cells
Lab Team 8 - Relaix
Street address 105 bd de l'Hôpital
City Paris
ZIP/Postal code 75013
Country France
Platform ID GPL1261
Series (1)
GSE65927 Early postnatal expression data from mouse skeletal muscle stem cells

Data table header descriptions
VALUE Data after processing by the Combat algorithm (log2)

Data table
1415670_at 9.37577848
1415671_at 8.838924142
1415672_at 9.800727727
1415673_at 6.664490309
1415674_a_at 8.131194732
1415675_at 8.583586078
1415676_a_at 10.08452463
1415677_at 7.729225092
1415678_at 10.09463626
1415679_at 9.946977542
1415680_at 9.131808326
1415681_at 8.523867099
1415682_at 8.051268542
1415683_at 9.952265621
1415684_at 6.225661303
1415685_at 7.70506094
1415686_at 9.726263527
1415687_a_at 9.278516161
1415688_at 9.341596644
1415689_s_at 8.441839369

Total number of rows: 45101

Table truncated, full table size 1026 Kbytes.

Supplementary file Size Download File type/resource
GSM1611273_P12_C.CEL.gz 3.7 Mb (ftp)(http) CEL
Raw data provided as supplementary file
Processed data included within Sample table

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