Transcriptome analysis of flower sex differentiation in Jatropha curcas L. using RNA sequencing
RNA samples from flower buds in six different developmental phases were pooled for a transcriptome analysis to obtain comprehensive gene expression information during the process of flower sexual differentiation. After DNase I treatment, magnetic beads with oligo (dT) were used to isolate mRNA. The mRNA was then fragmented into short fragments in fragmentation buffer, and cDNA was synthesized using the mRNA fragments as the template. The resulting short cDNA fragments were purified with a QIAquick PCR extraction kit and resolved in EB buffer. After the fragment ends were repaired and poly (A) tailed, the short fragments were ligated to sequencing adapters. Suitable fragments were selected as templates for PCR amplification. During the QC steps, an Agilent 2100 Bioanalyzer and ABI Step OnePlus Real-Time PCR System were used for the quantification and qualification of the sample library. Sequencing of the library was performed using an Illumina HiSeq™ 2000 at Beijing Genome Institute (Shenzhen, China).