RNA-seq of Ascoseira mirabilis
Total RNA was extracted using an RNAprep pure plant kit (Tiangen, Beijing, China) according to the manufacturers instructions, after which the concentration, quality and integrity were determined using a Nano Drop spectrophotometer (Thermo Scientific). Three micrograms of RNA were used as input material for the RNA sample preparations. Sequencing libraries were generated using the TruSeq RNA Sample Preparation Kit (Illumina, San Diego, CA, USA). After adenylation of the 3 ends of the DNA fragments, Illumina PE adapter oligo nucleotides were ligated to prepare for hybridization. To select cDNA fragments of the preferred 200 bp in length, the library fragments were purified using the AMPure XP system (Beckman Coulter,Beverly, CA, USA). DNA fragments with ligated adaptor molecules on both ends were selectively enriched using Illumina PCR Primer Cocktail in a 15 cycle PCR reaction. Products were purified (AMPure XP system) and quantified using the Agilent high sensitivity DNA assay on a Bioanalyzer 2100 system (Agilent). The sequencinglibrary was then sequenced on a Hiseq platform (Illumina) by Shanghai Personal Biotechnology Cp. Ltd.