Discovery of microRNAs Associated with Myogenesis by Deep Sequencing of Serial Developmental Skeletal Muscles in Pigs
18~30 nt RNA fragments were excised and purified from a PAGE gel, and adaptors were ligated to their 5’ and 3’ ends using T4 RNA ligase. RT-PCR was performed using adaptor primers for 17 cycles and the 90 bp DNA fragments were subsequently isolated from agarose gel. Subsequently, the purified DNA was used directly for sequencing analysis using the Illumina Genome Analyzer (Illumina) according to the manufacturer's instructions. The 35-nt sequence tags were obtained from Solexa sequencing.
cDNA