title: GSM8543153: Yeast, wildtype, 1h40m, rep2; Saccharomyces cerevisiae; RNA-Seq
library_construction_protocol: RNA was extracted from yeast cells using Qiagen RNeasy mini kit (Cat No.74106). The cell pellet was resuspended in buffer RLT and incubated at room temperature for 5 min to achieve complete lysis. The lysate mixed with half volume of absolute alcohol was loaded into RNeasy spin column placed in 2 ml collection tube. The tubes were centrifuge at 8,000 rpm for 1 min, and flow through discarded. On column DNase I (Cat No.79254) treatment and subsequent column washes was performed according to manufacturer's protocol. RNA was eluted from the column using Nuclease free water.The concentration and purity of RNA quantified using Nanodrop Spectrophotometer (Thermo Scientific; 2000). The integrity of RNA in the samples assessed on Tapestation (Agilent). RNA concentration quantified using Qubit RNA HS assay kit (Q32855). RNA sequencing libraries were prepared with Illumina-compatible NEBNext® Ultra™ II Directional RNA Library Prep Kit following manufacturer's protocol