radish transcriptome from NAU-LU127
Total RNA was isolated using Trizol reagents (Invitrogen, USA) according to the manufacturer’s protocol. For cDNA library construction, a total of 20 µg RNA from radish leaf samples at two different stages were pooled in equimolar quantity. Briefly, the mRNA was isolated using magnetic beads with Oligo (dT), and fragmented to small pieces using fragmentation buffer (Ambion, USA). Then the mRNA fragments were used as templates to synthesize double-stranded cDNA with random hexamer primers using the SuperScript Double-Stranded cDNA Synthesis Kit (Invitrogen, USA). The synthesized cDNA were purified with QiaQuick PCR extraction kit and subjected to end reparation and single nucleotide A (adenine) addition. Thereafter, the short fragments were connected with adapters and the suitable fragments were screened as templates for PCR amplification. The transcriptome library was sequenced using Illumina HiSeqTM 2000.
Nanjing agriculture university
radish