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The raw fast5 files produced by nanopore direct RNA sequcing. Three samples, Col0 and two mutants, and three biological replications of each sample were sequencing. Each biological replication only contains one run (fllow cell). Thus, tolal nine runs were produced.The mRNAs were isolated from about 75 ug total RNA of per-sample using the Dynabeads mRNA purification kit (Thermo Fisher Scientific) following the instructions to prepare the libraries for nanopore direct RNA sequencing. And the quality and quantity of mRNA was assessed though the NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific). Then the nanopore libraries were prepared from 1 μg poly(A)+ RNA with the direct RNA sequencing Kit (SQK-RNA001, Oxford Nanopore Technologies) according to the detailed manufacturer’s instructions. And the sequencing was realized the nanopore direct RNA sequencing using MinION MkIb with R9.4 flowcells. |
RNA-Seq |
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PolyA |
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