After filtering, expression matrices were written to file in tab-delimited format. Briefly, sample quality was verified by assessment of MA plots, normalized unscaled standard error (NUSE all samples had median log250 in at least 4 of the 23 samples. Raw data (.CEL Intensity files) were processed using R/Bioconductor packages. Healthy subject, primary myoblasts extracted from deltoid muscles and differentiated for 3 days to form myotubes The chips were scanned with the GeneChip® Scanner 3000 7G (Affymetrix) at a resolution of 0.7 µm. The chips were washed and stained in the GeneChip® Fluidics Station 450 (Affymetrix). The quality of RNA samples was assessed with Agilent 2100 Bioanalyzer.īiotinylated single strand cDNA targets were prepared, starting from 150 ng of total RNA, using the Ambion WT Expression Kit (Cat # 4411974) and the Affymetrix GeneChip® WT Terminal Labeling Kit (Cat # 900671) according to Affymetrix recommendations.įollowing fragmentation and end-labeling, 4.7 μg of cDNAs were hybridized for 16 hours at 45oC. Total RNA from muscle cells was extracted. Purified muscle stem cells were differentiated for 3 days into myotubes. Pantera has all but disowned their first four albums, this song is track 5 on the fourth of those albums, Power Metal.
After rinsing 3 times the proliferative myoblasts with PBS, and 3 times with DMEM to remove any FBS residual, the human muscle stem cells were differentiated into myotubes by culturing them in DMEM for 3 days. A minimum of 80% of the cell population were positive for desmin. The myogenic cell population was enriched using a CD56 magnetic bead and for their myogenicity using anti-desmin antibodies. Muscles biopsies were dissociated mechanically and plated in proliferation medium.
Muscle samples were obtained by open biopsy of deltoid muscle. GEO help: Mouse over screen elements for information.
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