GSM8014128
Tissues were directly lysed in 350 ul of RLT plus buffer with beta-mercaptoethanol from the Qiagen Rneasy Microkit (#74004). Tissue samples were subject to vortexing and stored at -80 C. For RNA extraction, tissue samples were thawed and homogenized using Qiashredder columns before processing with the Qiagen RNeasy Microkit following manufacturer instructions. According to instructions, a genomic DNA removal step was performed using genomic DNA removal columns. RNA was eluted in 16 ul of nuclease free water and RNA amounts were quantified using Qubit RNA-high sensitivity kit. 250 ng of total RNA was used for library construction. NEBNext Poly(A) mRNA Magnetic Isolation Module (#E7490L) was used for mRNA isolation according to manufacturer instructions. NEBNext® Ultra™ II Directional RNA Library Prep kit with Sample Purification Beads (#E7765L) was used for library preparation according to manufacturer instructions. Samples were sequenced using NovaSeq 6000. 100 bp pair-ended reads were obtained.