RNA-seq of Gal_1-2
A library was independently prepared with 1 g of total RNA for each sample using an Illumina TruSeq Stranded mRNA Sample Prep Kit (Illumina, Inc., San Diego, CA, USA, #RS-122-2101). The NEBNext rRNA Depletion Kit (NEB) was used to remove the rRNA in the total RNA, and the remaining mRNA was fragmented into small pieces using divalent cations under elevated temperatures. The cleaved RNA fragments were copied into the first-strand cDNA using SuperScript II reverse transcriptase (Invitrogen, #18064014) and random primers. This is followed by second-strand cDNA synthesis using DNA Polymerase I, RNase H, and dUTP. These cDNA fragments then went through an end repair process, adding a single A' base and ligating the adapters. The products were then purified and enriched with PCR to create the final cDNA library. The libraries were quantified using KAPA Library Quantification Kits for Illumina Sequencing platforms according to the qPCR Quantification Protocol Guide (KAPA BIOSYSTEMS, #KK4854) and assessed for quality using the TapeStation D1000 Screen Tape