RNA-seq of T449 (autotetraploid rice) ovary at pre-flowering stage, biological replicate 3
Total RNA was extracted from the hybridT449XH1 ovary by using Trizol reagent (Invitrogen, CA, USA) following the manufacturers procedure. The quantity and purity of total RNA were analysis by Bioanalyzer 2100 and RNA 6000 Nano LabChip Kit (Agilent, CA, USA) with RIN number > 7.0. Approximately 10 g of total RNA was used to deplete ribosomal RNA by using Epicentre Ribo-Zero Gold Kit (Illumina, San Diego, USA). Following purification, the poly(A)- or poly(A) + RNA fractions 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 Hiseq2500 at the (LC Sciences, USA) following the vendors recommended protocol.
SUB5302676
Ovary-F2-3