ONT Rapid Barcoding Kit, 3 x 96 reactions

The Rapid Sequencing Kits and Rapid Barcoding Kits V14 from Oxford Nanopore Technologies provide a quick and efficient method to prepare sequencing libraries from genomic DNA (gDNA). These kits are specifically designed for use on ONT devices such as MinION™, and they streamline workflows for sequencing native DNA while preserving base modifications.

Key Features

1. Rapid Sequencing of Native DNA

• Preserves Native Characteristics:
  • No PCR amplification required, eliminating PCR-induced bias.
  • Enables detection of base modifications (e.g., methylation) alongside the nucleotide sequence.
• Speed and Simplicity:
  • Library preparation in as little as 10 minutes for singleplex Rapid Sequencing Kits.
  • ~60 minutes for multiplex workflows using Rapid Barcoding Kits.
• Minimal Equipment:
  • Simplified process requiring basic laboratory equipment.

2. Multiplexing with Rapid Barcoding Kits

• Multiplex Up to 96 Samples:
  • Attach barcoded tags to template DNA to pool samples in a single sequencing run.
  • Useful for cases where individual sample data requirements are lower than the capacity of a flow cell.
• Efficient Use of Resources:
  • Reduces cost per sample and maximizes sequencing throughput by pooling multiple samples.
• Compatible Kits:
  • Rapid Barcoding Kit 24: For multiplexing up to 12 samples.
  • Rapid Barcoding Kit 96: For multiplexing up to 96 samples.

3. Workflow

1. Fragmentation and Tagging:
   • A transposase cleaves the DNA and simultaneously attaches tags to the cleaved ends.
2. Adapter Ligation:
   • Rapid Sequencing Adapters are ligated to the tagged DNA ends.
3. Barcoding (for Rapid Barcoding Kits):
   • Unique barcodes are attached to the DNA ends, allowing multiple samples to be pooled.
4. Pooling and Sequencing:
   • Pooled samples are loaded onto ONT flow cells for sequencing.
   • Barcoded reads are classified and sorted using MinKNOW software.

4. High Accuracy and Performance

• Chemistry Upgrade:
  • V14 chemistry is optimized for speed and simplicity, delivering improved accuracy and higher output with R10.4.1 Flow Cells.
• Q20+ Accuracy:
  • Capable of generating modal raw read accuracies of Q20+ (≥99%).
• Long Reads:
  • Long DNA fragments in the sample allow the generation of very long reads, despite transposase-induced fragmentation.

Kit Specifications

Compatibility

• Devices: MinION™, GridION™, and PromethION™.
• Flow Cells: R10.4.1 flow cells.

Applications

• Whole-genome sequencing.
• Epigenetic studies (e.g., methylation detection).
• Pathogen surveillance.
• Large-scale genomic studies with multiplexed samples.

Advantages

1. Time-Efficient:
   • Drastically reduces library preparation time compared to traditional methods.
2. Cost-Effective:
   • Multiplexing reduces cost per sample.
3. PCR-Free:
   • Retains DNA base modifications and eliminates amplification biases.
4. High Throughput:
   • Compatible with ONT devices for large-scale sequencing.

Shipping and Storage

• Shipped at 2–8°C.
• Store at -20°C upon receipt.
• Shelf life: 3 months from delivery.

Safety and Legal Note

• Not intended for health assessment, disease diagnosis, or treatment purposes.

The ONT Rapid Sequencing and Rapid Barcoding Kits V14 provide a user-friendly, efficient, and accurate approach to sequencing, catering to both small-scale and large-scale genomic studies. Their fast preparation time, multiplexing capabilities, and PCR-free workflows make them invaluable for modern sequencing applications.