Overview

Full-length adapters are compatible with ligation-based library construction workflows for direct and targeted multiplexed DNA and RNA (cDNA) sequencing applications on the Illumina® platform. Since sequencing barcodes are added during an adapter ligation step, full-length adapters are particularly well suited for PCR-free workflows.

KAPA Unique Dual-Indexed (UDI) Adapters have a standard Illumina adapter backbone. They are therefore easily incorporated in new and existing Illumina sequencing workflows, and do not require customized sequencing or indexing primers. A combination of 8-nt sequencing barcodes on each of the i5 and i7 adapter oligos supports paired-end sequencing on 2- and 4-channel Illumina instruments. KAPA UDI Adapters support a broad range of sample pooling (2- plex up to 96-plex, as appropriate for your workflow), for target enrichment and/or sequencing.

Unique dual-indexing is recommended for all applications and Illumina sequencers, whereas combinatorial dual-indexing may suffice for older generation instruments with non-patterned flow cells.1,2

Product highlights

 

Fewer misassigned reads improve confidence in results

  • Index misassignment during multiplexed sequencing may be the result of index hopping, barcode or sample cross-contamination, template switching during PCR amplification of pooled samples, and/or sequencing/analysis errors; some of which may be mitigated by adapter design and quality
  • Unique dual-index combinations in KAPA UDI Adapters allow reads with unexpected barcode combinations to be filtered out prior to data analysis
  • KAPA Adapters are manufactured using stringent procedures and undergo sequencing-based QC testing to reduce the potential for index misassignment resulting from barcode cross-contamination

 

High-performance, complete workflows for a wide range of applications

  • High-performance library construction workflows require appropriate, high-quality accessories such as adapters and beads to achieve optimal library yields and quality
  • KAPA Adapters undergo functional testing in an NGS library construction workflow to confirm high levels of library construction efficiency, minimal levels of adapter-dimer formation and consistent performance across all 96 index combinations

 

Flexible and efficient PCR-free human whole-genome sequencing

  • Even a small amount of library amplification may introduce bias which results in uneven coverage and the requirement for more sequencing; particularly when sequencing human and other complex genomes, or microbial genomes with extreme GC-content
  • PCR-free workflows have become the gold standard for human whole-genome sequencing (WGS)
  • High-quality, full-length KAPA Adapters used in combination with KAPA HyperPrep Kits support the flexible and highly efficient library construction protocols needed for high-throughput, PCR-free human WGS

 

  1. https://www.illumina.com/science/education/minimizing-index-hopping.html. Accessed February 2019.
  2. Costello M, et al. BMC Genomics. 2018;19:332