title: GSM5928748: h1_3_f3.rna; Arabidopsis thaliana; RNA-Seq
library_construction_protocol: Rosette leaves from the F3 generation were flash frozen in liquid nitrogen, pulverized with mortar and pestle on dry ice, and the resulting material was subjected to vortexing in Trizol (Invitrogen, cat. no. 15596-026).  Chloroform was then added at one-fifth the total volume and further vortexing was carried out until the solution appeared homogenous.  RNA was subsequently pelleted in ice-cold isopropanol. The resuspended RNA was subjected to rRNA removal with Ribo-zero plant kit (Illumina, MRZPL1224) according to the manufacturer's protocol. 50ng ribo-depleted was used for library preparation with the Scriptseq kit (Epicentre, cat. no. SSV21124) following the manufacturer's protocol but with the following modifications: the RNA fragmentation step was extended to 10 minutes, and the temperature was increased to 90°C.