title: GSM7247293: siMETTL3_input_rep1; Homo sapiens; RNA-Seq
library_construction_protocol: Input and polysome RNA samples were made to 150 μL in dH2O and cleaned by adding 280 μL Beckman RNAClean XP beads. Samples were mixed by pipetting up and down, incubated at room temperature for 5 minutes, pelleted on a magnetic rack for 5 minutes, washed with 1000 μL 70% EtOH three times, and air dried for ten minutes before eluting into 150 μL dH2O. Poly(A)-selection of cleaned RNA samples was then done using Promega PolyATract mRNA Isolation System IV per the manufacturer's protocol. Briefly, RNA samples were made to 500 μL and incubated at 65°C for 10 minutes before addition of 3 μL Biotin-dT and 1x SSC. Once cooled, this sample was added to 100 μL of washed poly(A) magnetic beads in 0.5x SSC. Following a 10-minute incubation at room temperature, RNA-bound beads were captured using a magnetic rack, washed in 0.1X SSC, and eluted in a total of 250 μL dH2O. RNA libraries for Nanopore direct RNA seq were prepared with Oxford Nanopore technology SQK-RNA-002 kit following manufacturer's protocols.