title: Carthamus tinctorius L. flower unigenes
organism: Carthamus tinctorius
description: In brief, beads with oligo(dT) were used to isolate poly(A) mRNA after total mixed RNA was collected from flower tissue. The fragmentation buffer was added for interrupting mRNA to short fragments; and then taking these short fragments as templates, random hexamer-primer was used to synthesize the first-strand cDNA. The second-strand cDNA was synthesized using buffer, dNTPs, RNase H and DNA polymerase I respectively. The paired-end library was synthesized using the Genomic Sample Prep kit (Illumina, San Diego, CA) according to the manufacturer’s instructions. Short fragments were purified with the QiaQuick PCR extraction kit and then resolved with EB buffer for end repair and adding poly (A). After that, the short fragments were connected with sequencing adapters and then suitable fragments were separated by agarose gel electrophoresis. Finally, the sequencing library was built by amplification with PCR and the library was sequenced using Illumina HiSeq™ 2000 platform.