GSM9340298: AML12 cells, Control4; Mus musculus; RNA-Seq
Invitrogen TRIZOL Reagent were used for RNA extraction; VAHTS Universal V6 RNA-seq Library Prep Kit for Illumina were used for library construction (1) The Nanodrop2000 microspectrophotometer detects sample purity and concentration; (2) The Labchip GX touch microfluidic capillary system detects the integrity of RNA samples. (1) The library was constructed starting with Total RNA and mRNA with ployA tails was enriched by Oligo(dT) magnetic beads; (2) Add Fragmentation Buffer for mRNA fragmentation; (3) One strand of cDNA was synthesised using a six-base random primer using mRNA as a template, followed by the synthesis of the second strand of cDNA using buffer, dNTPs, DNA polymerase I and RNase H, and cDNA purification; (4) Purified double-stranded cDNA was subjected to end repair, base A was added, sequencing junctions were added, and fragments were screened to recover cDNA around 350 bp; (5) PCR enrichment was performed to obtain cDNA libraries; (6) Double-stranded cDNA is denatured at high temperature into single-stranded DNA, and cyclisation primers are added to form a single stranded Circular DNA library (single stranded Circular DNA).