RNA seq of Ammopiptanthus nanus: leaf
Total RNA was extracted using TRIzol? reagent (Invitrogen, Burlington, Canada) according to the manufacturerĄŻs protocol. The quality and purity of RNA samples were assessed by Bioanalyzer 2100 (Agilent Technologies, Santa Clara, USA) and RNA 6000 Nano LabChip Kit (Agilent, CA, USA) with RIN number >7.0. Magnetic beads with Oligo (dT) (Invitrogen, Burlington, Canada) were used to isolate mRNA from 10 ug total RNA. Following purification, the mRNA was cleaved into short fragments with divalent cation under elevated temperature. Then the cleaved RNA fragments were reverse transcribed to create the final cDNA library in accordance with the protocol for the mRNA-Seq sample preparation kit (Illumina, San Diego, USA), the average insert size for the paired-end libraries was 300 bp (ĄŔ50bp). And then we performed the paired-end sequencing on an Illumina Hiseq2000 at the (LC Sceiences, USA) following the vendorĄŻs protocol.