title: GSM3592217: Gfp_4; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was extracted using Trizol Reagent precisely per manufacturer protocol. Library preparation was performed with 10ug of total RNA with a Bioanalyzer RIN score greater than 8.0. Ribosomal RNA was removed by poly-A selection using Oligo-dT beads (mRNA Direct kit, Life Technologies). mRNA was then fragmented in buffer containing 40mM Tris Acetate pH 8.2, 100mM Potassium Acetate and 30mM Magnesium Acetate and heating to 94 degrees for 150 seconds. mRNA was reverse transcribed to yield cDNA using SuperScript III RT enzyme (Life Technologies, per manufacturer's instructions) and random hexamers. A second strand reaction was performed to yield ds-cDNA. cDNA was blunt ended, had an A base added to the 3' ends, and then had Illumina sequencing adapters ligated to the ends. Ligated fragments were then amplified for 12 cycles using primers incorporating unique index tags.  Fragments were sequenced on an Illumina HiSeq-3000 using single reads extending 50 bases.
attributes.tag: GEO Accession
attributes.value: GSM3592217