title: Lotus cornioulatus Transcriptome via Illumina Second Generation Sequen
design: Beads with Oligo(dT) are used to isolate poly(A) mRNA after total RNA is collected from eukaryote (prokaryocyte can be treated with kit to remove rRNA before next step). Fragmentation buffer is added for interrupting mRNA to short fragments. Taking these short fragments as templates, random hexamer-primer is used to synthesize the first-strand cDNA. The second-strand cDNA is synthesized using buffer, dNTPs, RNaseH and DNA polymerase I, respectively. Short fragments are purified with QiaQuick PCR extraction kit and resolved with EB buffer for end reparation and adding poly(A). After that, the short fragments are connected with sequencing adapters. And, after the agarose gel electrophoresis, the suitable fragment are selected for the PCR amplification as templates. At last, the library could be sequenced using Illumina HiSeq™ 2000.