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RNA sequencing.

Mk Mohamad Al kadi
EI Eiji Ishii
DT Dang Tat Truong
DM Daisuke Motooka
SM Shigeaki Matsuda
TI Tetsuya Iida
TK Toshio Kodama
DO Daisuke Okuzaki
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Illumina libraries were prepared using a TruSeq stranded total RNA kit (Illumina, USA) following the manufacturer’s instructions and sequenced on a HiSeq 3000 sequencer (Illumina).

A direct RNA sequencing library was prepared using a direct RNA sequencing kit (SQK-RNA001; Oxford Nanopore Technologies) from approximately 500 ng of poly(A) RNA input following the manufacturer’s instructions. Briefly, a reverse transcription adapter was ligated using T4 DNA ligase (New England Biolabs), and first-strand cDNA was synthesized using SuperScript III reverse transcriptase (Thermo Fisher Scientific). Sequencing adapters were ligated using T4 DNA ligase (New England Biolabs). The library was loaded onto a MinION flow cell and sequenced for 48 h. The RNA calibration strand (RCS) spike-in was added to the run in the first step as a control.

Copyright and License information:  ©2021 Al kadi et al.
This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.

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