Transcriptome sequencing of 02428-2x (diploid rice) anthers, the anthers at the meiosis stage
The quantity and purity of total RNA were analysis by Bioanalyzer 2100 and RNA 6000 Nano LabChip Kit (Agilent, CA, USA) with RIN number > 8. Approximately 10 ug of total RNA representing a specific adipose type was subjected to isolate Poly (A) mRNA with poly-T oligo attached magnetic beads (Invitrogen, CA, USA). Following purification, the mRNA is fragmented into small pieces using divalent cations under elevated temperature. Then the cleaved RNA fragments were reverse-transcribed to construct the final cDNA library according to the manuscript of the mRNA-Seq sample preparation kit (Illumina, San Diego, USA). Finally, we performed the paired-end sequencing with the average insert size of 300 bp (±50 bp) on an Illumina Hiseq2000 (LC Sciences, USA) following the vendor’s recommended protocol.