title: GSM8548501: 7h_stimulus_biorep2; Arabidopsis thaliana; RNA-Seq
library_name: GSM8548501
library_construction_protocol: Total RNA was extracted using the Quick-RNA Plant mini-prep kit (Zymo) and Treated with DNase I. Total RNA samples (100-500 ng) were hybridized with Ribo-Zero Plant to substantially deplete cytoplasmic, mitochondrial and chlorpoplast rRNA from the samples.  Stranded RNA sequencing libraries were prepared as described using the Illumina TruSeq Stranded Total RNA Kit with Ribo-Zero Gold (RS-122-2401 and RS-122-2402).  Purified libraries were qualified on an Agilent Technologies 2200 TapeStation using a D1000 ScreenTape assay (cat# 5067-5582 and 5067-5583).  The molarity of adapter-modified molecules was defined by quantitative PCR using the Kapa Biosystems Kapa Library Quant Kit (cat#KK4824).  Individual libraries were normalized to 10 nM and equal volumes were pooled in preparation for Illumina sequence analysis. NovaSeq 150x150 bp Sequencing:  Sequencing libraries were chemically denatured and applied to an Illumina NovaSeq flow cell using the NovaSeq XP workflow (20043131).  Following transfer of the flowcell to an Illumina NovaSeq 6000 instrument, a 151 x 151 cycle paired end sequence run was performed using a NovaSeq 6000 S4 reagent Kit v1.5 (20028312).