S16-S17-S18
Approximately 1 g of RNA was used for 3' mRNA-Seq library construction using the QuantSeq 3' mRNA-Seq Library Prep Kit FWD for Illumina. To minimize lane effects, the samples were multiplexed, using the multiplexing sequencing adaptors provided in the Multiplexing Sample Preparation Oligo Kit (Illumina). Size selection was performed on a 2% agarose gel (Low Range Ultra Agarose, Biorad 161-3107). The denatured library was diluted to a final concentration of 6 pM and loaded on a flow cell (Illumina). After cluster generation, the multiplexed library was sequenced on an Illumina NextSeq500 (75 cycles, single end, high output).