Direct RNA sequencing (SQK-RNA004) library preparation
Direct RNA nanopore sequencing library preparation using the SQK-RNA004 kit for native RNA strand analysis.
Version History
Version 1.0 Current
Effective: 2026-02-22Procedure Details
Equipment (Catalog)
7
-
Microcentrifuge
Centrifugation
Microcentrifuge for brief spins -
Micropipette
Liquid handling
Micropipettes -
Magnetic rack
Magnetic separation
Magnetic rack for RNAClean XP bead cleanup -
Fluorometer
Measurement
Qubit fluorometer for library QC -
Vortex mixer
Mixing
Vortex mixer -
Nanopore sequencer
Sequencing
MinION / GridION / PromethION sequencer -
Thermal cycler
Thermal regulation
Thermal cycler for RT and adapter ligation
Materials (Catalog)
11
-
Ethanol
Chemical
70% ethanol for RNAClean XP bead washes -
Microcentrifuge tube
Consumable
1.5 mL Eppendorf DNA LoBind tubes -
Pipette tips
Consumable
Wide-bore pipette tips for RNA handling -
T4 DNA Ligase (2,000,000 U/mL)
(NEB)
M0202T/MDNA ligase
T4 DNA Ligase (M0202T/M) -
SuperScript III Reverse Transcriptase
(Thermo Fisher)
18080044Reverse transcriptase
SuperScript III Reverse Transcriptase (18080044) -
Direct RNA Sequencing Kit
(Oxford Nanopore)
SQK-RNA004Sequencing kit
Direct RNA Sequencing Kit (SQK-RNA004) -
10 mM dNTP Solution Mix
(NEB)
N0447dNTP mix
10 mM dNTP Solution Mix (N0447) -
RNaseOUT Recombinant Ribonuclease Inhibitor
(Invitrogen)
10777019Enzyme mix
RNaseOUT Ribonuclease Inhibitor (10777019) -
NEBNext Quick Ligation Reaction Buffer
(NEB)
B6058Ligation reagent
NEBNext Quick Ligation Reaction Buffer (B6058) -
Agencourt RNAClean XP
(Beckman Coulter)
A63987Magnetic beads (SPRI)
Agencourt RNAClean XP beads (A63987) -
Nuclease-free water
Reagent
For elution
Procedure Steps (Version 1.0)
No steps defined for this version.
References
- Oxford Nanopore Technologies — Direct RNA Sequencing (SQK-RNA004) Protocol, Rev B, 20 Sep 2023 Link protocol