title: Illumina HiSeq 2500 paired end sequencing; Tumor invasion in draining lymph nodes is associated with Treg accumulation in breast cancer patients
design: Tumor invasion in draining lymph nodes is associated with Treg accumulation in breast cancer patients
library_construction_protocol: CD4+CD45RA-CD25- (Memory CD4+ Tconv), CD4+CD45RA-CD25high (Memory Treg), were sorted by flow cytometry in a BD FACS ARIA II cell sorter, with a purity of 98-99%. Cells were collected in TCL buffer (Qiagen) with 1% of b-mercaptoethanol and stored at -80 until subsequent analysis.  RNA was isolated using a Single Cell RNA purification kit (Norgen),including RNase-Free DNase Set (Qiagen) treatment. RNA integrity number was evaluated with an Agilent RNA 600s0 pico kit.  Retrotranscription was carry out with SMARTv4 low input kit (Takara). Barcoded Illumina compatible libraries were generated from 5-10ng of DNA of each sample, using Nextera XTP reparation Kit.
attributes.tag: Experimental Factor: sampling site
attributes.value: primary tumour
attributes.tag: Experimental Factor: cell type
attributes.value: alpha-beta T cell