title: GSM1695162: circRNA_identification_RBC; Homo sapiens; RNA-Seq
library_construction_protocol: Leukocyte depleted Red Blood Cells, from an infection free donor with A+ blood type,  was obtained from the Scottish National Blood Transfusion Service (surplus stock, stored for 27 days post transfusion in red cell storage solution). Leukodepletion filtration reduced white blood cells to <1.85x10E4 per ml, and plasma was removed before resuspension in storage solution. RBCs were pelleted and snap frozen on dry ice prior to RNA extraction using Trizol (Life technologies) and treatment with DNase I (Promega) according to the manufacturer’s instructions. RNA quantitation was performed using the NanoDrop ND-1000 spectrophotometer (NanoDrop) and quality was assessed using the Agilent 2100 Bioanalyser (Agilent Technologies). 400ng of RNA was used in library preparation and sequencing. Library was prepared based on Illumina's guidelines for TruSeq RiboZero stranded mRNA LT sample prep kit, following the alternative fragmentation protocol. Paired-end reads were sequenced using a paired end 100 base pair protocol on an Illumina HiSeq2000.
attributes.tag: GEO Accession
attributes.value: GSM1695162