RNA-Seq of Eimeria stiedai: FL-cDNA
The isolated RNAs from the Ou, Os, Mz4, and Gc samples were pooled in equal amounts to provide the total RNA for PacBio SMRT sequencing. The mRNA was isolated from the total RNA using magneticoligo-dTbeads and reversely transcribed into cDNA using Clontech SMARTer PCR cDNA Synthesis Kit (Clontech Laboratories, Inc. CA, USA). Size selection of the PCR products was performed using BluePippin Size Selection System (Sage Science, Beverly, MA, USA), and the fragments ranging from 0.5 to 6 kb were retained. Afterwards, large-scale PCR was carried out to amplify the full-length cDNA. The ends of the cDNAs were repaired before the sequencing adapters were ligated to the cDNAs. SMRT-bell template libraries were constructed from the obtained cDNAs and sequenced on the PacBio Sequel platform.