Nature research paper: mRNA ageing shapes the Cap2 methylome in mammalian mRNA
A549 cells were seeded in a single well of a six-well plate. The following day, cells were transfected with either 2.5 µg FTSJD1 double nickase plasmid or a mixture of 1.5 µg FTSJD1 and 1.5 µg RIG-I double nickase plasmids using Lipofectamine 3000 transfection reagent according to the manufacturer’s instructions. The procedure for isolation of single-cell KO clones was performed as described for HEK293T cells.
. Forty-eight-hour-old zebrafish embryos were collected in TRIzol reagent and stored at −80 °C until further use.Total RNA was isolated from cells using TRIzol reagent according to the manufacturer’s instructions unless otherwise stated. For isolation of total RNA from mouse tissues, TRIzol-submerged tissues were homogenized with high-impact zirconium 1.5-mm beads twice at 50 Hz for 3 min using TissueLyser II .
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