mix-2_HJM3FALXX_L5
After the RNA sample was tested, the eukaryotic mRNA was enriched with magnetic beads with Oligo (dT). Subsequently, the fragmentation buffer was used to break the mRNA into short fragments, and mRNA was used as a template to synthesize a strand cDNA with random hexamers. Then, buffer, dNTPs (dTTP in dNTP was replaced with dUTP) and DNA Polymerase were added. I and RNase H synthesize the double-stranded cDNA, and then purify the double-stranded cDNA with AMPure XP beads, and then degrade the second strand of the cDNA containing U with USER enzyme. The purified double-stranded cDNA was firstly end-repaired, A-tailed and ligated to the sequencing linker, and the fragment size was selected using AMPure XP beads. Finally, PCR amplification was performed and the PCR product was purified using AMPure XP beads to obtain the final library.
SUB5912839
RRB06734-w