miRNA-Seq of mus musculus: tumor from IL-21R deficiency (IL-21RKO) male mouse with hepatocellular carcinoma (HCC) at 27 weeks of age
The RNA was sequentially ligated with 3 and 5 adaptors. Thereafter, reverse transcription reaction was applied and the PCR products were purified with PAGE gel. The purified PCR products were subsequently denatured into single-stranded DNA for circularization, and were further replicated via rolling cycle amplification, resulting in a DNA nanoball (DNB) containing multiple copies of DNA. Sufficient quality DNBs were subsequently loaded into patterned nanoarrays by using high-intensity DNA nanochip technique and sequenced through combinatorial Probe-Anchor Synthesis (cPAS).