Detection P values <0.01 were used to filter all data. Significant ?2 differential expression called on Log2 transformed data with an adjusted P value if <0.005
Array probe summaries were calculated in Beadstudio and quantile normalised
Labelled with Ambion Total Prep RNA amplification Kit.
Thymocytes were isolated from wt and mutant animals, red blood cells lysed and stained for TCRb and CD5 cell surface antigens and the TCRb/CD5low populations flow sorted, pelleted and stored at -80 degrees prior to total RNA extraction.
Mice were housed in standard clean conditions and were used at 2-3 weeks of age
Standard Trizol reagent extraction
(Parameters: Extracted product = total_RNA, Amplification = none)
Scanned in Illumina BeadArray Reader using BeadStudio
Hybridised at 58 C for 16 hours in oven with rocking platform