GSM2236272: Sample_WT_3; Citrus trifoliata; RNA-Seq
Total RNA was isolated from the WT and the MT (three biological replicates per genotype) using the Plant RNAiso Plus according to the manufacturer’s instructions (Takara, Kusatsu, Japan). Genomic DNA was removed from total RNA by DNase treatment (Promega, Madison, WI, USA). All RNA samples were quantified and examined for protein contamination and reagent contamination by a Nanodrop ND 1000 spectrophotometer (NanoDrop, Wilmington, DE, USA). Due to some lncRNAs lacking the poly(A) tail, total RNA was treated to remove rRNA by using Ribo-Zero rRNA Removal Kits (Plant) according to the manufacturer’s instructions (illumina, USA), retaining lncRNA both with and without a poly(A) tail. Six strand-specific RNA libraries with an insert size of about 250–500 nucleotides were prepared according to a UTP method (Parkhomchuk et al., 2009), and submitted to the Beijing Genomics Institute (BGI, Shenzhen, China) for 125 bp paired-end sequencing on the Illumina HiSeq 2500, at a depth of approximately 90 million reads per library.