gender: male age: 31 years cell type: platelet time: stored for 8 days
Treatment protocol
Apheresis platelets were stored at 22±2 ℃ in storage tank and 10ml sample were collected at 2 day, 5 day, 8 day. The 10 ml platelet product was centrifuged at 1500g for 10 minutes to removal of the plasma. The harvested samples were snap-frozen in liquid nitrogen until being complete frozen , then stored at -80 ℃ until processing.
Extracted molecule
total RNA
Extraction protocol
Total RNA was isolated from apheresis platelet using mirVanaTM RNA Isolation Kit in accordance with the manufacturer’s protocol.
Label
biotin
Label protocol
Total RNA were amplified, labeled and purified by using GeneChip WT 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
Gene expression in platelets stored for 8 day A8.CEL
Data processing
Raw data were normalized by RMA algorithm, Gene Spring Software 13.1 (Agilent technologies, Santa Clara, CA, US).
