C57BL/10ScSnJ mice were used to have SP cells of TA muscle
Extracted molecule
total RNA
Extraction protocol
Total RNA was isolated using RNeasy micro kit (Qiagen, Valencia, CA).
Label
Biotin
Label protocol
RNAs were subjected to linear amplification using the GeneChip® Two cycle target-labeling protocol (Affymetrix Inc., Santa Clara, CA, USA).
Hybridization protocol
The GeneChip® Two cycle target-labeling protocol (Affymetrix Inc., Santa Clara, CA, USA).
Scan protocol
Agilent model G2500A GeneArray scanner used.
Description
SP cell preparations were washed, pelleted and resuspended in 20µl of PBS and stored at –80oC until processing for microarray analysis using manufacturers instructions and previously described methods from our laboratory . Total RNA was isolated using RNeasy micro kit (Qiagen, Valencia, CA). The purity, concentration and integrity of RNA was determined using a Nanodrop ND-1000 spectrophotometer (NanoDrop Technologies, Wilmington, DE) and an Agilent 2100 Bioanalyzer (Agilent Technologies Inc., Palo Alto, CA). RNAs were subjected to linear amplification using the GeneChip® Two cycle target-labeling protocol (Affymetrix Inc., Santa Clara, CA, USA). Six independently separated EOM and six TA SP cell preparations were used for microarray analysis using the Affymetrix® Mouse 430 ver 2.0 GeneChip arrays. Raw intensities for each probe set were stored in electronic formats by expression summaries calculated using Microarray Suite version 5.0 (MAS5) algorithms.