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ONT Ligation Sequencing Kit V14 SQK-LSK114, 6 Reactions
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Specifications:
| Application | Next Generation Sequencing | ||
| Storage Temperature | 2-8°C | ||
| Product Type | Sequencing Reagents | Forms | Kit with Various components |
| Product Brand | Oxford Nanopore Technology | ||
| Product Grade | Molecular Biology | ||
High-Accuracy, Flexible Library Preparation for Long-Read Sequencing Applications
The ONT Ligation Sequencing Kit V14 (SQK-LSK114) provides a versatile and robust method for preparing sequencing libraries from double-stranded DNA (dsDNA), including genomic DNA (gDNA), amplicons, and cDNA. Designed to work seamlessly with ONT’s R10.4.1 nanopores, this kit enables users to achieve Q20+ modal raw read accuracy (≥99%), high throughput, and long-read capabilities.
Ideal for applications requiring control over read length, amplification-free workflows, and preservation of native base modifications, SQK-LSK114 is compatible with upstream processes such as whole genome amplification (WGA), target enrichment, and size selection using the Short Fragment Eliminator kit (EXP-SFE001).
Key Features & Benefits
-
Q20+ Read Accuracy
- Optimized for use with R10.4.1 nanopore for modal raw read accuracy ≥99%
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Flexible Sample Input
- Accepts 1 µg gDNA or 100–200 fmol amplicons/cDNA
-
Read Length = Fragment Length
- Preserve full-length reads by using high molecular weight (HMW) DNA
- Optional fragmentation for smaller insert workflows
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Rapid & Reliable Workflow
- Total prep time: ~60 minutes
- No PCR required (optional for low-input samples)
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Long-Run Optimization
- Includes fuel fix and improved adapter chemistry for extended run durations
-
Multiplexing Options
- Compatible with Native Barcoding Kits (24 and 96), PCR Barcoding Kits, and Multiplex Ligation Kit XL
Applications
- Whole genome sequencing (WGS)
- Human and microbial genome analysis
- Target enrichment and RRMS
- Metagenomics
- Transcriptomics (direct or PCR cDNA)
- Cell-free DNA (cfDNA) and single-cell studies
- Structural variant detection
- Methylation and epigenetic profiling
Library Preparation Overview
- End repair/dA-tailing using NEBNext Companion Module
- Adapter ligation using V14 chemistry
- Magnetic bead purification
- Optional size selection for long-read optimization
Kit Contents (6 Reactions)
| Component | Cap Color | Fill Volume (µL) | Qty |
|---|---|---|---|
| DNA Control Sample (DCS, 3.6 kb) | Yellow | 35 | 1 vial |
| Ligation Adapter (LA) | Green | 40 | 1 vial |
| AMPure XP Beads | Amber | 1200 | 1 vial |
| Ligation Buffer (LNB) | White | 200 | 1 vial |
| Long Fragment Buffer (LFB) | Orange | 1800 | 2 vials |
| Short Fragment Buffer (SFB) | Clear | 1800 | 2 vials |
| Sequencing Buffer (SB) | Red | 700 | 1 vial |
| Elution Buffer (EB) | Black | 1200 | 1 vial |
| Library Beads (LIB) | Pink | 600 | 1 vial |
| Library Solution (LIS) | White w/ pink label | 600 | 1 vial |
| Flow Cell Flush (FCF) | Clear/Blue | 8000 | 1 vial |
| Flow Cell Tether (FCT) | Purple | 200 | 1 vial |
AMPure XP Reagent (Beckman Coulter) included and stable at -20°C.
Performance Specifications
| Feature | Specification |
|---|---|
| Prep Time | ~60 minutes |
| Input Requirement | 1 µg gDNA or 100–200 fmol amplicons/cDNA |
| PCR Required | No (optional for low input) |
| Fragmentation | Optional; not recommended for long-read workflows |
| Read Length | Equal to input fragment size |
| Chemistry | Kit 14 (V14) |
| Storage | -20°C (long-term), 2–8°C (shipping) |
| Shelf Life | 3 months from receipt |
Required 3rd Party Materials
- NEBNext Companion Module v2 (E7672S or E7672L)
- DNA LoBind tubes, PCR tubes, nuclease-free water
- 80% ethanol (freshly prepared)
- Qubit™ dsDNA HS Assay Kit + Qubit Fluorometer
- Magnetic rack, thermal cycler, microcentrifuge, pipettes
Compatible Devices & Flow Cells
| Devices | MinION Mk1B, Mk1C, Mk1D, GridION, PromethION, Flongle |
|---|---|
| Flow Cells | FLO-MIN114, FLO-PRO114M, FLO-FLG114 |
Associated Protocols
- Whole genome sequencing (WGS)
- Amplicon sequencing
- Direct cDNA sequencing
- Single-cell transcriptomics (10X Genomics, 3'/5')
- Cell-free DNA and methylation studies
- Low input workflows with PCR
Ordering Information
| Catalog No. | Description | Pack Size |
|---|---|---|
| SQK-LSK114 | Ligation Sequencing Kit V14 | 6 reactions |
For large-scale workflows, refer to Multiplex Ligation Kit XL V14 (SQK-MLK114.96-XL)
The ONT Ligation Sequencing Kit V14 (SQK-LSK114) offers unmatched flexibility, speed, and accuracy for nanopore sequencing applications. Optimized for long reads, native DNA, and methylation analysis, it is the kit of choice for researchers seeking Q20+ basecalling accuracy and robust data across diverse genomic applications.