NEBNext® Companion Module for Oxford Nanopore Technologies® Direct RNA Sequencing

Product Information

The NEBNext Companion Module for Oxford Nanopore Technologies Direct RNA Sequencing is a convenient solution that simplifies direct RNA library preparation. It includes the ONT-validated NEB ligation and reverse transcription reagents for use with the ONT SQK-RNA004 protocol (non-barcoded workflow), provided in volumes optimized for this workflow.

Included Reagents:

  • Induro Reverse Transcriptase
  • Induro RT Reaction Buffer
  • Deoxynucleotide (dNTP) Solution Mix
  • NEBNext Quick Ligation Reaction Buffer
  • T4 DNA Ligase
  • RNase Inhibitor, Murine
  • Nuclease-free Water

All reagents are manufactured to stringent quality standards. Each lot is performance tested as a complete set through direct RNA library preparation and sequencing on an ONT device to support consistent results.

This module is used in conjunction with the ONT Direct RNA Sequencing Kit (SQK-RNA004/-XL). The appropriate device-specific SQK-RNA004 protocol from ONT should be followed for use of these reagents.

Figure 1: The NEBNext Companion Module for Oxford Nanopore Technologies Direct RNA Sequencing simplifies direct RNA library preparation
NEBNext Companion Module for Oxford Nanopore Direct RNA Sequencing workflow

The NEBNext Companion Module contains the NEB reagents recommended in the SQK RNA004 workflow and is used with the Oxford Nanopore Technologies Direct RNA Sequencing Kit (SQK RNA004/-XL) to prepare non-barcoded direct RNA sequencing libraries from poly(A)-tailed or total RNA. In this workflow, the RT Adapter (RTA) is ligated to poly(A) RNA, followed by reverse transcription to generate a complementary cDNA strand that stabilizes the RNA and improves sequencing output. The resulting RNA–cDNA hybrid is then ligated to the RNA Ligation Adapter (RLA) for loading and sequencing on Oxford Nanopore Technologies instruments.

This product is related to the following categories:

Library Preparation for Oxford Nanopore Technologies Next Generation Sequencing Library Preparation

Product Information

The NEBNext Companion Module for Oxford Nanopore Technologies Direct RNA Sequencing is a convenient solution that simplifies direct RNA library preparation. It includes the ONT-validated NEB ligation and reverse transcription reagents for use with the ONT SQK-RNA004 protocol (non-barcoded workflow), provided in volumes optimized for this workflow.

Included Reagents:

  • Induro Reverse Transcriptase
  • Induro RT Reaction Buffer
  • Deoxynucleotide (dNTP) Solution Mix
  • NEBNext Quick Ligation Reaction Buffer
  • T4 DNA Ligase
  • RNase Inhibitor, Murine
  • Nuclease-free Water

All reagents are manufactured to stringent quality standards. Each lot is performance tested as a complete set through direct RNA library preparation and sequencing on an ONT device to support consistent results.

This module is used in conjunction with the ONT Direct RNA Sequencing Kit (SQK-RNA004/-XL). The appropriate device-specific SQK-RNA004 protocol from ONT should be followed for use of these reagents.

Figure 1: The NEBNext Companion Module for Oxford Nanopore Technologies Direct RNA Sequencing simplifies direct RNA library preparation
NEBNext Companion Module for Oxford Nanopore Direct RNA Sequencing workflow

The NEBNext Companion Module contains the NEB reagents recommended in the SQK RNA004 workflow and is used with the Oxford Nanopore Technologies Direct RNA Sequencing Kit (SQK RNA004/-XL) to prepare non-barcoded direct RNA sequencing libraries from poly(A)-tailed or total RNA. In this workflow, the RT Adapter (RTA) is ligated to poly(A) RNA, followed by reverse transcription to generate a complementary cDNA strand that stabilizes the RNA and improves sequencing output. The resulting RNA–cDNA hybrid is then ligated to the RNA Ligation Adapter (RLA) for loading and sequencing on Oxford Nanopore Technologies instruments.

This product is related to the following categories:

Library Preparation for Oxford Nanopore Technologies Next Generation Sequencing Library Preparation

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
E3440S Induro® Reverse Transcriptase E3437AVIAL -20 1 x 0.048 ml 200,000 units/ml
Induro® RT Reaction Buffer E3438AVIAL -20 1 x 0.192 ml 5 X
Deoxynucleotide (dNTP) Solution Mix E3439AVIAL -20 1 x 0.048 ml 10 mM
NEBNext® Quick Ligation Reaction Buffer E3441AVIAL -20 1 x 0.264 ml 5 X
T4 DNA Ligase E3442AVIAL -20 1 x 0.108 ml 2,000,000 units/ml
RNase Inhibitor, Murine E3443AVIAL -20 1 x 0.024 ml 40,000 units/ml
Nuclease-free Water E3444AVIAL -20 1 x 1.1 ml Not Applicable

Related Products

Companion Products

NEBNext® High Input Poly(A) mRNA Isolation Module
Monarch® Spin RNA Isolation Kit (Mini)
NEBNext® Poly(A) mRNA Magnetic Isolation Module
NEBNext® Magnetic Separation Rack

Protocols, Manuals & Usage

Usage Guidelines

Usage guidelines for reagents included in the NEBNext® Companion Module for Oxford Nanopore Technologies® Direct RNA Sequencing (NEB #E3440)

FAQs & Troubleshooting

FAQs

1. Which protocol should be followed when using the NEBNext® Companion Module for Oxford Nanopore Technologies® Direct RNA Sequencing (NEB #E3440)?
2. Are the reagents supplied in the NEBNext® Companion Module for Oxford Nanopore Technologies® Direct RNA Sequencing (NEB #E3440) sufficient to construct libraries for Oxford Nanopore Technologies devices?
3. Can the reagents supplied in the NEBNext® Companion Module for Oxford Nanopore Technologies® Direct RNA Sequencing (NEB #E3440) be used for multiplex / barcoded sequencing (SQK-DRB004.24 workflow)?

Citations & Technical Literature

Citations

Product Citation Tool

Product Citation Tool

Bioz-powered product citation information for E3440.

Quality, Safety & Legal

Quality Control Assays

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specifications & Change Notifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

E3440S_v1

Certificate of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

E3440S_v1_10341850
E3440S_v1_10356770

Legal and Disclaimers