Blueberry Transcriptome - fruit skins
Total RNA from the skin and pulp was isolated by BGI using the TRIzol® Reagent (Invitrogen). RNA quality was characterized by the quotient of the 28S to 18S ribosomal RNA electropherogram peak using a Nanodrop 2000 instrument (Thermo Scientific) and a Bioanalyzer Chip RNA 7500 series II (Agilent). Two libraries were prepared using the mRNA-Seq 8 sample prep Kit (Illumina) according to the manufacturer's instructions. First, magnetic beads containing poly-T molecules were used to isolate poly(A) mRNA from 20 µg of total RNA. Then, samples were fragmented by fragmentation buffer and reverse transcribed into cDNA with a PrimeScriptTM 1st Strand cDNA Synthesis Kit (TaKaRa). Next, end repair and A-base tailing was purified using the QIAquick PCR Purification Kit (QIAGEN). Finally, Illumina adapters were ligated to the cDNA fragments. Each library had an insert size of 200 bp, and 75 bp sequences were generated on an Illumina Genome Analyzer IIx platform.