RNA-seq of kiwifruit flowers
Total RNAs were isolated from young floral buds using Trizol Reagent. mRNA was purified with the RNA Clean-up Kit according to the manufacturerĄŻs instructions. The purified RNA was subsequently fragmented into ~200 bp pieces by treatment with divalent cations at 75 ĄăC. The first strand cDNA synthesis was generated from the cleaved RNA fragments using reverse transcriptase (Invitrogen) with random hexamer primers, and second-strand cDNA was synthesized using RNase H and DNA polymerase I (New England BioLabs). Second generation sequencing was performed on the Illumina HiSeq 2000 instrument in Shanghai Haiyu Biotechnology Co., Ltd.