Native barcoding expansion

Dodávateľ: Oxford Nanopore Technologies
Native barcoding expansion
Činidla pro nukleové kyseliny Next Generation Sequencing Reagents
Native Barcoding Expansion packs provide a versatile method of preparing barcoded nanopore sequencing libraries for applications where high throughput is required.

  • Multiplex samples to reduce price per sample
  • PCR-free multiplexing — preserve and analyse additional information such as base modifications
  • High sequencing yields
  • Control over read length
  • Suitable for upstream processes such as size selection or whole-genome amplification

Barcoding or multiplexing is useful when the amount of data required per sample is less than the total amount of data that can be generated from a single flow cell: it allows a user to pool multiple samples and sequence them together, making more efficient use of the flow-cell. Multiplexing samples onto one flow cell can reduce the cost per sample

Native Barcoding Expansion 1-12 provides 12 unique barcodes with sufficient reagents for six sequencing libraries.
Native Barcoding Expansion 13-24 provides 12 unique barcodes (different to those provided in the Native Barcoding Expansion 1-12) with sufficient reagents for six sequencing libraries.
Native Barcoding Expansion 96 provides 96 unique barcodes with sufficient reagents for 12 sequencing libraries.

These kits enable PCR-free multiplexing of dsDNA samples such as gDNA – use in combination with Ligation Sequencing Kit and cDNA – use in combination with Direct cDNA Sequencing Kit

The library preparation method is similar to the Ligation Sequencing Kit protocol; DNA ends are repaired and dA-tailed using the NEBNext End Repair/dA-tailing module, and then a unique dT-tailed barcode adapter is ligated on the dA-tailed template. Barcoded samples are then pooled together. Each barcode adapter also has a cohesive end and this is used as a hook to ligate to the supplied sequencing adapters.

These kits are optimized to generate maximum throughput, without the need for PCR. For highest data yields, we recommend starting with a total of 200 fmol of pure input DNA (for two samples, this means starting with 100 fmol of each). Starting with lower amounts of input material or impure samples can affect library preparation efficiency and lower sequencing throughput.

Native Barcoding Expansion packs are compatible with upstream processes such as whole genome amplification (for applications where under 1 ng of sample is available) and size selection (for enrichment of specified fragment lengths, using the BluePippin, for example). When size selecting, we recommend increasing the amount of input used, as size selection can be a wasteful process.

Deconvolution of barcoded sequencing data is supported by the MinKNOW and Guppy software, which classify the barcode sequence and sort reads into corresponding folders.

Native Barcoding Expansion packs can be used together with: Ligation Sequencing Kit (76487-138); Direct cDNA Sequencing Kit (76487-162); Flow Cell Wash Kit (76487-116).

Sample multiplexing for Native Barcoding Expansion 1-12 and Native Barcoding Expansion 13-24 is ≤12, while Native Barcoding Expansion 96 is ≤96.

Objednávacie informácie: These products are anticipated to be discontinued in September 2023. We recommend using the Native Barcoding Kits V14, which utilise the latest chemistry and R10.4.1 Flow Cells for enhanced sequence accuracy.

Informace o dodávce: Flow cells and kits are shipped together at 2 to 8°C. Upon receipt, please place the product in a long-term storage location at –20°C. Oxford Nanopore Technologies deem the useful life of the product to be 3 months from receipt by the customer.

Pozor: Oxford Nanopore Technologies products are not intended for use for health assessment or to diagnose, treat, mitigate, cure, or prevent any disease or condition.
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