title: Leaf transcriptomic of Paphiopedilum hirsutissimum
design: The mature leaves cDNA library of Paphiopedilum hirsutissimum was prepared using Illumina’s kit (Illumina, San Diego, CA, USA) following manufacturer’s recommendations. Briefly, the poly(A) mRNA was purified from 6.0 µg total RNA of mature leaves of P. hirsutissimum using oligo(dT) magnetic beads and fragmented into short sequences using divalent cations under elevated temperature. First and second strand cDNA were synthesized from cleaved RNA fragments. After the end repair and ligation of adaptors, the products were cleaned up with a QIAquick PCR Purification Kit (Qiagen, Valencia, CA) to create the final cDNA library. Finally, after validating on an Agilent Technologies 2100 Bioanalyzer using the Agilent DNA 1000 chip kit, the mature leaves cDNA library of P. hirsutissimum was sequenced using Illumina HiSeqTM 2000 platform to obtain short sequences from both ends at Shanghai Biotechnology Corporation (SBC) in Shanghai, China.