NEBNext® Flu A Integrated Indexing Primer Module
NEBNext® Flu A Integrated Indexing Primer Module
NEBNext Flu A Integrated Indexing Primer Module is intended for use in cDNA synthesis, amplification, and library preparation upstream of Influenza A (Flu A) sequencing. Libraries generated with the Primer Module are meant for sequencing with the Oxford Nanopore Technologies® (ONT) Native Barcoding workflow.
- Compatible with various Influenza A strains (e.g., H3N2, H1N1, H5N1) from various hosts (e.g., Human, Avian, Bovine)
- Integrated indexing enables pooling of up to 48 samples following RT-PCR
- Primers incorporate Oxford Nanopore Native Barcoding sequences
- Universal Influenza A targeting reduces the impact of variants on amplification efficiency
| Catalog # | Concentration | Size |
|---|---|---|
| E3436S | Not Applicable | 48 reactions |
Product Information
For laboratories interested in sequencing the Influenza A genome, streamlined and specific cDNA synthesis is an essential first step. The NEBNext Flu A Integrated Indexing Primer Module is a curated set of 48 plated primer pairs for use in RT-PCR upstream of NGS library preparation. Downstream, the libraries prepared are compatible with ONT Native Barcoding.
The positioning of the integrated indices in the primer sequences decreases the risk for variants to impact amplification efficiency. Refer to the product manual for the full list of required materials not supplied.
The NEBNext Flu A Integrated Indexing Primer Module has undergone extensive QCs to ensure that the primer pairs are balanced, well-aligned with clinically relevant Flu A sequences, and free of cross contamination. In-house testing includes the full recommended workflow, combining RT-PCR, NGS library prep, and sequencing followed by analysis of sequencing metrics.
Due to its modular nature, the NEBNext Flu A Integrated Indexing Primer Module provides flexibility for maximal ease of use. Full information on the primer sequences included can be found at the NEBNext Flu A primer sequence repository.
Note: The workflow can also support sequencing libraries prepared with the NEBNext Flu A Integrated Indexing Primer Module on Illumina® sequencers as described in the relevant FAQs.
The NEBNext Flu A Integrated Indexing Primer Module is designed to be paired with LunaScript® Multiplex One-Step RT-PCR Kit for streamlined cDNA synthesis and targeted amplification, followed by library preparation with Oxford Nanopore Technologies Ligation Sequencing Kit (SQK-LSK114).
The optional use of these Flu A cDNA amplicons as unindexed inputs for Illumina library prep is also supported using the NEBNext UltraExpress® FS DNA Library Prep Kit (NEB #E3340).
This product is related to the following categories
Reagents Supplied
The following reagents are supplied with this product:
| Component Name | Component # | Stored at (°C) | Amount | Concentration |
|---|---|---|---|---|
| NEBNext® Flu A Integrated Indexing Primer Module | E3436AVIAL | -20 | 1 × 5 μl/well | Not Applicable |
Related Products
Companion Products
- LunaScript® Multiplex One-Step RT-PCR Kit
- NEBNext® dA-Tailing Module
- NEBNext® Quick Ligation Module
- Monarch® Mag Viral DNA/RNA Extraction Kit
Protocols, Manuals & Usage
Protocols for this product
- Where can I find protocols for use with the NEBNext Flu A Integrated Indexing Primer Module (NEB #E3436)?
Manuals
The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls.
- manualE3436
Application Notes
- Facilitating Purification and Detection of Viral Nucleic Acids from Milk
- A Sample to Results Workflow for Influenza A Sequencing
FAQs & Troubleshooting
FAQs
- Which additional reagents are needed for cDNA synthesis and amplification with the NEBNext Flu A Integrated Indexing Primer Module (NEB #E3436)?
- Where can I find NEBNext Flu A Integrated Indexing Primer (NEB #E3436) sequences?
- Which Flu A strains can be targeted with the NEBNext Flu A Integrated Indexing Primers (NEB #E3436)?
- Which library prep reagents and protocols can be used for sequencing cDNA amplicons made with the NEBNext Flu A Integrated Indexing Primer Module (NEB #E3436)?
- Is there a recommended Ct value-range for Flu A inputs when using the NEBNext Flu A Integrated Primer Module (NEB #E3436)?
- What is the minimum read depth needed for Flu A sequencing when using the NEBNext Flu A Integrated Primer Module (NEB #E3436)?
- What sample types can the NEBNext Flu A Integrated Indexing Primer Module (NEB #E3436) be used with?
- What extraction methods are recommended for use in the NEBNext Flu A sequencing workflow when using the NEBNext Flu A Integrated Primer Module (NEB #E3436)?
- The ONT SQK-LSK114 kits include AMPure® XP Beads, but are additional beads needed to run the NEBNext Flu A Integrated Indexing Primer Module (NEB #E3436) ONT® library prep workflow?
