title: GSM1213287: Cytoplasmic small RNA preparation; Homo sapiens; RNA-Seq
library_construction_protocol: Subcellular fractionation was carried out essentially as in Bai and Laiho (Efficient sequential recovery of nucleolar macromolecular components. Proteomics, 12, 3044-3048, 2012). Total RNA was extracted from whole cells and the isolated cellular compartments using the TRIzol. Approximately 400 µg RNA from each compartment was used to prepare cDNA libraries. Small RNAs (less than 200 nt) were isolated from the total RNA samples using mirVana Isolation Kit and separated on denaturing 15% polyacrylamide gel (15% PAA, 19:1 acrylamide/bis, 7 M urea) to purify RNAs in the approximate range of 10 - 40 nt.  The sRNA size range was confirmed and the RNA was quantified using Bioanalyzer with Agilent Small RNA kit. cDNA libraries were constructed using the Ion Torrent RNA-seq kit v1 for small RNA libraries, and their purity and concentration was confirmed using Bioanalyzer with the Agilent DNA kit. sRNA library sequencing was performed on a 314 chip on a Ion Torrent sequencing platform.
attributes.tag: GEO Accession
attributes.value: GSM1213287