GSM8791819: un-damaged cartilage3; Homo sapiens; ncRNA-Seq
Firstly, ribosomal RNA is removed from the total RNA, and then RNase R enzyme is used to break the RNA into short fragments of 250-300 bp. The fragmented RNA is used as a template, and random oligonucleotides are used as primers to synthesize the first strand of cDNA. Subsequently, RNase H is used to degrade the RNA strand, and dNTPs (dUTP, dATP, dGTP, and dCTP) are used as raw materials to synthesize the second strand of cDNA in the DNA polymerase I system. The purified double stranded cDNA was subjected to end repair, followed by the addition of an A-stail and sequencing adapter connection. The cDNA with a size of around 350-400 bp was screened using AMPure XP beads. Use USER enzyme to degrade the second strand of cDNA containing U, and finally perform PCR amplification to obtain a library.