RNA-Seq of Atox1 knockdown
After the total RNA samples were tested and qualified, they were purified according to the characteristics of mRNA from different sources. Eukaryotic mRNA with polyA tail structure at the 3' end was used for enrichment and purification.polyA tail structure at the 3' end of eukaryotic mRNAs, magnetic beads with Oligo (dT) were selected for enrichment and purification; prokaryotic mRNAs did not have this property, and the rRNA was removed with the kit to obtain more valid information.Prokaryotic mRNAs do not have this characteristic, so the rRNA was removed with the kit to obtain more effective information. Fragmentation Buer was added to the purified mRNA to fragment it into short fragments.Fragmentation Buer was added to the purified mRNA to fragment it into short fragments. The fragmented RNA was then used as a template for reverse transcription using random primers to realize the first strand synthesis of cDNA, and the 2nd Strand was added.The second strand of cDNA was synthesized by adding 2nd Strand Marking Buer and 2nd Strand/End Repair Enzyme Mix. The second strand of cDNA was synthesized by adding 2nd Strand Marking Buer andEnd Repair, add base A, add sequencing connector, recover the target fragment by magnetic bead screening, and PCR amplification, thus completing the whole library preparation.library preparation