tissue: peripheral blood cell type: lymphocytes disease status: healthy control
Growth protocol
Venous peripheral blood from fasting patients and healthy controls were collected during 7am to 9am.Peripheral blood lymphocytes were separated by Ficoll gradient centrifugation using Ficoll-PlaqueTM Plus (GE, Sweden) according to the manufacturer's protocol.
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted from lymphocytes using Trizol reagent (Invitrogen) according to the manufacturer's protocol. RNA concentrations were determined by Nanodrop ND-1000 (Nanodrop Technologies, Wilmington, DE). RNA quality was assessed on an Agilent 2100 Bioanalyzer
Label
biotin
Label protocol
Total RNA were amplified, labeled and purified by using GeneChip 3’IVT Express Kit (Cat#901229, Affymetrix, Santa Clara, CA, US) followed the manufacturer’s instructions to obtain biotin labeled cRNA.
Hybridization protocol
Array hybridization and wash was performed using GeneChip® Hybridization, Wash and Stain Kit (Cat#900720, Affymetrix, Santa Clara, CA, US)in Hybridization Oven 645 (Cat#00-0331-220V, Affymetrix, Santa Clara, CA, US)and Fluidics Station 450 (Cat#00-0079, Affymetrix, Santa Clara, CA, US) followed the manufacturer’s instructions.
Scan protocol
Slides were scanned by GeneChip® Scanner 3000 (Cat#00-00212, Affymetrix, Santa Clara, CA, US) and Command Console Software 3.1 (Affymetrix, Santa Clara, CA, US) with default settings.
Description
CB2007224-C4.CEL health control 4
Data processing
Raw data were normalized by RMA method, Gene Spring Software 11.0 (Agilent technologies, Santa Clara, CA, US). The values were log2 transformed.
