title: RNA-Seq of Oryza Sativa: indica cultivated rice leaf tissue
design: Collected leaf tissue was frozen at -80oC and ground with mortar and pestles. A Qiagen RNeasy Plant Mini Kit (Valencia,CA) was used to extract RNA. A Bioanalyzer 2100 (Agilent Technologies, Inc.) was used to qualify and quantify RNA samples. Only RNA samples with RNA Integrity Number (RIN) > 7.5 were used for library preparation. Libraries were constructed with NEBNext mRNA Library Prep Master Mix Set for Illumina kit (New England BioLabs Inc.). Total RNA was fragmented into smaller pieces, and then synthesized into the first and second strand of cDNA with reverse transcriptae and random primers. NEBNext singleplex Oligos for Illumina as indexes of each library were added during the preparation (New England BioLabs Inc).