title: GSM2476147: D-2 ESC; Mus musculus; Bisulfite-Seq
library_construction_protocol: generation protocol: Dusp9 cDNA was cloned into the shuttle plasmid pBS31 using NotI and MfeI digestion. The resulting plasmid was electroporated into KH2 ESCs together with a plasmid driving expression of Flp recombinase (Beard et al., 2006). Genomic DNA was collected using phenol/chloroform extraction RRBS libraries were prepared using a commercial kit (Ovation RRBS Methyl-Seq System, NuGen, San Carlos, CA) following the manufacturer's protocol except that we pooled 12 individually barcoded reactions after the final repair step and performed the bisulfite conversion and library amplification as a pool. Libraries were sequenced on an Illumina HiSeq2500 high-output flowcell without a PhiX spike-in using Nugen's custom sequencing primer for read 1 (36 bases) and standard Illumina sequencing primers to read the 8-base sample barcodes.
attributes.tag: GEO Accession
attributes.value: GSM2476147