GSM6858440: RNA_Mycelia_H153G_DMSO2; Colletotrichum orbiculare; RNA-Seq
Cells were filtered, scraped, and soaked in liquid nitrogen with 600 µl of lysis buffer or 400 µl of Trizol reagent. The frozen cells and lysis buffer grains were milled by a Multi-beads Shocker at 2,800 rpm for 15 sec for 1 cycle. For ribosome profiling, RNase I treatment, ribosome pelleting, size selection, preadenylated linker ligation, rRNA depeletion, reverse transcription, cDNA circularization, and PCR amplification. For RNA-Seq, Poly(A) selection and cDNA synthesis were performed using Illumina Stranded mRNA Prep, Ligation (Illumina) and subsequent steps were performed by TruSeq Stranded Total RNA Library Prep Gold (Illumina).