GSM5929901: 35S::CBP60g, 28C, DC3000 Rep 2; Arabidopsis thaliana; RNA-Seq
20 to 60 mg of fresh leaf tissues were flash-frozen in liquid nitrogen and ground using a TissueLyser (Qiagen). Plant RNA was extracted using a Qiagen Plant RNeasy Mini Kit following the manufacturer's protocol, including the recommended on-column DNase I digestion. Libraries were prepared using the Illumina TruSeq Stranded mRNA Library Preparation Kit on a Perkin Elmer Sciclone G3 robot following manufacturer's recommendations. Completed libraries were QC'd and quantified using a combination of Qubit dsDNA HS and Advanced Analytical Fragment Analyzer High Sensitivity DNA NGS assays. The libraries were divided into two pools of 12 libraries each, combining them in equimolar proportions for multiplexed sequencing. Pools were quantified using the Kapa Biosystems Illumina Library Quantification qPCR kit. Each pool was loaded onto one (1) lane of an Illumina HiSeq 4000 flow cell and sequencing performed in a 1x50bp single read format using HiSeq 4000 SBS reagents. Base calling was done by Illumina Real Time Analysis (RTA) v2.7.7 and output of RTA was demultiplexed and converted to FastQ format with Illumina Bcl2fastq v2.19.1.