title: GSM1860686: M.27/1-8/C; Malus domestica; miRNA-Seq
library_construction_protocol: Leaf samples were ground with a mortar and pestle and chilled with liquid nitrogen. The frozen powder was mixed with extraction buffer (2% cetyltrimethylammonium bromide, 2% polyvinylpyrolidone, 100 mMTris pH 8.0, 25 mMethylenediaminetetraacetic acid, 2 M NaCl, 0.5 g L-1 (w/v) spermidine, 2% ß-mercaptoethanol) and total RNA was isolated using 0.8 M LiCl and isopropanol (Sigma).  Total RNA concentration and purity were determined using NanoDrop (Thermo Fischer Scientific). Total RNA quality was determined withBioAnalyzer (Agilent). The RNA samples had high and consistent RNA integrity values. 18 apple libraries were prepared with TruSeq Small RNA Sample Preparation Kit (Illumina) according to the manufacturers protocol. 50 nt single-end sequencing on a HiSeq2500 (Illumina) in rapid run mode was performed in the Genomics Core Facility of the Pennsylvania State Univeristy at University Park, PA. Adapter sequences were removed with Short Stack v.1.2.4 Redundant and nonredundant reads in each library were counted and RPM (reads per million) values were calculated
attributes.tag: GEO Accession
attributes.value: GSM1860686