title: Illumina HiSeq 2000 sinlge-end Sequencing of KAS21008422 (KAE21087950)
design: After removal of adapter sequences, all the trimmed reads were clustered according to the unique base sequence of the reads using CD-HIT (http://weizhong-lab.ucsd.edu/cd-hit/). To identify miRNA sequences, unique sequences identified after clustering were mapped to known miRNAs in the miRBase database (http://www.mirbase.org/) by blast search.
library_construction_protocol: Total RNA was extracted using TRIzol reagent according to the manufacturer's protocol Invitrogen, USA. Extracted RNA was purified using RNeasy Mini Kit Columns Qiagen, USA. MiRNA was isolated using a High Pure miRNA Isolation Kit Roche, USA. The 5' and 3' ends of the total RNAs were ligated with adapters. Subsequently, the cDNAs synthesized from the ligated RNAs were amplified by PCR. The enriched PCR products were separated by PAGE, and the small fragments approximately 120??40 bp were isolated from the gel. These fragments were used for cluster generation and sequencing on an Illumina HiSeq 2000 instrument Illumina, Inc., USA.