ncRNA-Seq of mus musculus: healthy liver from wild type (WT) male mouse fed on normal chow (NC) diet for nine weeks
The RNA was treated with RNase H kit to remove rRNA, following RNA fragmentation reaction. Thereafter, the first-strand and the second-strand cDNAs were sequentially synthesized. Next, double-stranded cDNA fragments were subjected to end-repair, and then a single A nucleotide was added to the 3 ends of the blunt fragments. The adaptors were subsequently ligated to the cDNAs, followed by PCR amplification. The PCR products were 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).