RNA-Seq of wheat ZM16
Total RNA was extracted from leaves using Trizol. mRNA was purified with Oligo(dT)-attached magnetic beads and broken into short fragments. Then, first-strand cDNA was generated using random hexamer-primed reverse transcription, followed by second-strand cDNA synthesis. The cDNA fragments were amplified by PCR, and the products were purified by Ampure XP Beads and validated using the Agilent Technologies 2100 bioanalyzer for quality control. The double stranded PCR were heated denatured and circularized by the splint oligo sequence to get the final library. The final library was amplified with phi29 to make DNA nanoballs (DNBs) with more than 300 copies in each molecule, DNBs were loaded into a patterned nanoarray, and 100-base paired-end reads were sequenced using the BGIseq500 platform