GSM7729986: Control_6h_3 [circRNA]; Mus musculus; RNA-Seq
Liver tissue of C57 BL6 mice were treated with liquid nitrogen immediately and maintained at -80C. 1 ug of total RNA was used for the construction of sequencing libraries. Ribosomal RNA is extracted from the total RNA, which is subsequently fragmented into short sequences ranging from 250-300 bp. Utilizing this fragmented RNA as a template and employing random oligonucleotides as primers, the first strand of cDNA is synthesized. Following this, the RNA strand is degraded using RNase H. The second strand of cDNA is then synthesized using dNTPs (dUTP, dATP, dGTP, and dCTP) within the DNA polymerase I system. The purified double-stranded cDNA undergoes processes such as end repair, A-tailed addition, and sequencing linkages. AMPure XP beads are employed to isolate cDNA fragments approximately 350-400 bp in length. The USER enzyme is used to degrade the second cDNA strand containing U, culminating in PCR amplification to produce the final library.