strain: Smarta cell type: day 60 memory PSGL1hi CD4 T Cell
Extracted molecule
total RNA
Extraction protocol
RNA was isolated with RNeasy mini elute kit (QIAGEN, Valencia, CA)
Label
biotin
Label protocol
Double stranded cDNA and biotin-labeled cRNA was synthesized and purified from 500 ng of total RNA. Purification of the cRNA and integrity of the cRNA was be assessed by running aliquots on the Bioanalyzer prior to hybridization. Hybridization buffer from the BeadChip kit (Illumina) was mixed with 1500 ng of biotin-labeled cRNA, heated to 65°C for 5 minutes, and then loaded onto the BeadChip. The BeadChips were sealed in a hybridization chamber and placed in an oven at 58°C with a rocker for 16-20 hours. After the hybridization, the BeadChips were washed and stained in a series of washes and stains as outlined in the Illumina protocol. The BeadChips were scanned on the Illumina Iscan.
Hybridization protocol
Standard Illumina hybridization protocol
Scan protocol
Standard Illumina scanning protocol
Description
CD4 T Cell
Data processing
The data were normalised using quantile normalisation with lumi in R
