ONT Rapid Sequencing Kit V14

The ONT Rapid Sequencing Kit V14 is a PCR-free sequencing library preparation kit that simplifies and speeds up the process of DNA sequencing. It provides an efficient and easy-to-use workflow for nucleic acid analysis with minimal sample preparation time. The kit employs transposase-based DNA fragmentation, which cleaves the DNA template and adds sequencing adapters for real-time nanopore sequencing.

Key Features:

1. Short Preparation Time:
   • 10-minute preparation time, ideal for users who require fast sequencing with minimal hands-on time.
2. No PCR Requirement:
   • The kit does not require PCR amplification, eliminating PCR bias and preserving the original base modifications.
3. Efficient DNA Fragmentation:
   • Transposase cleaves template DNA and attaches adapters, allowing for efficient DNA fragmentation and rapid library preparation.
4. Modular Configuration:
   • The kit uses the latest Kit 14 chemistry, which improves modal raw read sequencing accuracies and provides higher output on the latest nanopore R10.4.1.
5. Long Reads with High Accuracy:
   • Capable of producing long reads while maintaining high sequencing accuracy (over 99% accuracy (Q20+)).
6. Minimal Sample Manipulation:
   • Requires minimal pipetting and no clean-up steps, which simplifies the workflow and reduces potential contamination.
7. Higher Efficiency:
   • The kit is optimized for samples containing long fragments (greater than 30 kb) to achieve better sequencing output and longer read lengths.
8. Control of Fragmentation and Adapter Addition:
   • The use of Rapid Sequencing Adapters ensures clean and efficient sequencing with no extra preparation steps.
9. Flexible Sequencing:
   • Compatible with multiple flow cells such as FLO-MIN114 and FLO-FLG114, and compatible with various devices like Flongle, MinION Mk1B, MinION Mk1C, and PromethION.

Kit Components:

Additional 3rd Party Materials:

• Consumables:
  • 1.5 mL Eppendorf DNA LoBind tubes
  • 0.2 mL thin-walled PCR tubes
  • Nuclease-free water (e.g., ThermoFisher, Cat # AM9937)
  • Optional: Bovine Serum Albumin (BSA) (50 mg/ml) (e.g., Invitrogen™ UltraPure™ BSA, Cat# AM2616)
• Equipment:
  • Microfuge
  • Pipettes: P1000, P100, P20, P10, P2
  • Timer
  • Thermal cycler or heat block at 30°C and 80°C
• Optional Equipment:
  • Qubit fluorometer (or equivalent for QC checks)

Compatibility:

• Flow Cells:
  • FLO-MIN114, FLO-PRO114M, FLO-FLG114
• Devices:
  • Flongle, MinION Mk1B, MinION Mk1C, MinION Mk1D, GridION, PromethION

Applications:

1. Genomics:
   • Suitable for genomic sequencing and variant detection in various biological samples.
2. Microbial Sequencing:
   • Ideal for metagenomic studies, microbial community profiling, and microbial species identification.
3. Epigenetic Studies:
   • The PCR-free nature of the kit ensures preservation of base modifications for epigenetic analysis.
4. Transcriptomics:
   • Use in RNA sequencing to study gene expression and splicing events in various organisms.
5. Environmental and Agricultural Research:
   • Used for studying environmental microbiomes or agricultural samples, such as plant roots, soil microbiomes, and soil health monitoring.

Storage and Handling:

• Storage Temperature: Store the kit at −20°C for long-term storage.
• Handling: Thaw reagents at room temperature before use. Follow protocol guidelines for optimal performance.

The ONT Rapid Sequencing Kit V14 offers fast, simple, and efficient sequencing with minimal sample manipulation. Designed for high throughput and long-read sequencing, this kit ensures high accuracy for applications in genomics, microbial sequencing, and environmental studies.