title: RNAseq of Claviceps (Cp20.1) infected Secale cereale: young ovaries (5 dpi): Sample Rye infected with Claviceps purpurea strain Cp20.1
design: Beads with Oligo(dT) were used to isolate poly(A) mRNA after total RNA was collected from infected rye ovaries Fragmentation buffer is added for interrupting mRNA to short fragments. 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, RNaseH and DNA polymerase I, respectively. Short fragments were purified with QiaQuick PCR extraction kit and resolved with EB buffer for end reparation and tailing A. After that, the short fragments were connected with sequencing adapters. And, after the agarose gel electrophoresis, the suitable fragment were selected for the PCR amplification as templates. At last, the library was sequenced using Illumina HiSeq™ 2000.