Mononuclear cells (MNCs) were isolated, and then stored at -80 ℃ before RNA extraction.
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted using RNAiso (TaKaRa, Dalian, China).
Label
biotin
Label protocol
The sample labeling, microarray hybridization and washing were performed based on the manufacturer’s standard protocols. Briefly, total RNAs were transcribed to double strand cDNAs and then synthesized cRNAs. Next, 2nd cycle cDNAs were synthesized from cRNAs, followed by fragmentation and biotin labeling.
Hybridization protocol
The sample labeling, microarray hybridization and washing were performed based on the manufacturer’s standard protocols. Briefly, total RNAs were transcribed to double strand cDNAs and then synthesized cRNAs. Next, 2nd cycle cDNAs were synthesized from cRNAs. Followed fragmentation and biotin labeling, the 2nd cycle cDNAs were hybridized onto the microarray. After washing and staining, the arrays were scanned by the Affymetrix Scanner 3000(Affymetrix).
Scan protocol
Scanned by the Affymetrix Scanner 3000 (Affymetrix).
Description
HC-08
Data processing
Affymetrix GeneChip Command Console (version 4.0, Affymetrix) software was used to extract raw data. Next, Expression Console (version1.3.1, Affymetrix) software offered RMA normalization for both gene and exon level analysis. Then the gene expression analysis and alternative splice analysis proceeded separately.
