organism: Taxus mairei
description: Leaf was collected from a cultivated Taxus mairei of two years old. Total RNA was isolated with TRIzol (Invitrogen) according to the manufacturer's instructions. It was treated with RNase-free DNase I for 30 min at 37*C (New England BioLabs) to remove residual DNA.  Beads with oligo(dT) were used to isolate poly(A) mRNA after total RNA was collected from Taxus tissues. Fragmentation buffer was added for interrupting mRNA to short fragments. Taking these short fragments as templates, Random hexamer-primer were used to synthesize the first-strand cDNA. The second-strand cDNA was synthesized using buffer, dNTPs, RNase H and DNA polymerase I, respectively. Short fragments were purified with QiaQuick PCR extraction kit and resolved with EB buffer for end repair and adding poly(A). After that, the short fragments were connected with sequencing adapters. For amplification with PCR, we selected suitable fragments as templates, with respect to the result of agarose gel electrophoresis. At last, the library was sequenced using Illumina GA IIX.