library_construction_protocol: RNAs were extracted by Rneasy Mini kit The sequencing library is prepared by random fragmentation of the DNA or cDNA sample, followed  by 5' and 3' adapter ligation. Alternatively, "tagmentation" combines the fragmentation and ligation  reactions into a single step that greatly increases the efficiency of the library preparation process.  Adapter-ligated fragments are then PCR amplified and gel purified. For cluster generation, the library is loaded into a flow cell where fragments are captured on a lawn  of surface-bound oligos complementary to the library adapters. Each fragment is then amplified  into distinct, clonal clusters through bridge amplification. When cluster generation is complete, the  templates are ready for sequencing.