2.5 RNA preparation and construction of nanopore sequencing
library
Total RNA was isolated using the FastPure Universal Plant Total RNA
Isolation Kit (Vazyme, Nanjing, China) according to the manufacturer’s
instructions. RNA was cleaned and concentrated by NEB Next Poly(A) mRNA
Magnetic Isolation Module (E7490S) according to the manufacturer’s
instructions. Each nanopore DRS and transcriptome sequencing sample
contained 20 μg total RNA. Prepared RNA of each sample was used for a
nanopore DRS library preparation using the Oxford Nanopore DRS protocol
(SQK-RNA002, Oxford Nanopore Technologies) (Yahara et al., 2021). This
work was done in collaboration with Wuhan Benagen Technology Co, Ltd,
China.