title: GSM1480828: RNA WT Treg rep1; Mus musculus; RNA-Seq
library_construction_protocol: RNA-seq: Total RNA wasprepared from approximately 1 million cells by using mirVana miRNA Isolation Kit(AM1560, ABI). 200 ng of total RNA was subsequently used to prepare RNA-seq library byusing TruSeq SRRNA sample prep kit (FC-122-1001, Illumina) by following manufacturer’s6protocol.The resulted libraries are sequenced using the Illumina HiSeq 2000. Only unique reads of 50 bp were mapped to the mouse genome (mm9) using Tophat and Cufflinks. ChIP-seq: Cells were chemically cross linked by 1% formaldehyde. Cell lysates were made by appropriate sonication and protein-DNA complexes were isolated with antibody.  Libraries were prepared according to Illumina's instructions. Briefly, DNA was end-repaired and the blunt, phosphorylated ends were treated with Taq polymerase and dATP to yield a protruding 3- 'A' base for ligation of Illumina's adapters which have a single 'T' base overhang at the 3’ end. After adapter ligation, DNA was PCR amplified with Illumina primers for 15 cycles and library fragments of ~250 bp (insert plus adaptor and PCR primer sequences) were band isolated from an agarose gel. The purified DNA was captured on an Illumina flow celland sequenced on HiSeq 2000.
attributes.tag: GEO Accession
attributes.value: GSM1480828