GSM8968323: C57BL/6_17W_1; Mus musculus; miRNA-Seq
RNA was harvested using TRIzol (Invitrogen, Carlsbad, California, USA). 2 ug of total RNA was used for the construction of sequencing libraries. The library was constructed using the MGIEasy MiRNA Library Prep Kit (BGI-Shenzhen). RNA was sequentially ligated with 3' and 5' adapters, followed by reverse transcription into cDNA and amplification via PCR. Fragment size selection was performed using polyacrylamide gel electrophoresis to ensure quality control. The single-stranded library was then denatured and circularized, with uncyclized linear DNA molecules removed. The circularized DNA was amplified through phi29 and rolling circle amplification (RCA) to generate DNA nanoballs (DNBs), each containing over 300 copies of the original single-stranded circularized molecule. DNA nanoballs (DNBs) were deposited onto a patterned nanoarray, and paired-end 50-base sequencing reads were generated using the G400 platform (BGI-Shenzhen, China).