Accurate molecule counting is essential for somatic oncology applications, especially from low inputs of cfDNA where every molecule counts. This accuracy is enhanced by the use of Unique Molecular Identifiers (UMIs) incorporated into NGS adapters, providing barcodes for individual molecules.
The use of unique dual-indexing strategies can improve data quality by enabling the removal of reads containing unexpected barcode combinations, such as those caused by index hopping or index mis-assignment, prior to data analysis (Figure 3).
Such index mis-assignment can occur during several steps of multiplexed sequencing, including library preparation (cross-contamination of barcodes or samples), amplification of pooled samples (template switching during PCR), and/or errors during sequencing and analysis—especially when patterned flow cells are used for sequencing.
KAPA Adapters are a collection of high quality, unique dual-indexed adapters for ligation-based library construction. KAPA Adapters support many levels of sequencing multiplexing (2-plex up to 384-plex) on both two- and four-channel Illumina instruments that use either patterned or non-patterned flow cells.
(1)The sets of 192 and 768 barcode sequences used in KAPA UDI Adapters and KAPA UDI Primer Mixes (respectively) are exclusive to Roche and different between the two sets.
(2) Theoretically possible but not fully tested.
(3) This is sufficient for four or one library preps (KAPA UDI Adapter plate or KAPA UDI Primer Mixes plate, respectively) with the KAPA HyperPrep or KAPA HyperPlus Kit if no adapter dilution is required. Generous overfill supports use on automated liquid handling systems.
(4)KAPA UDI Primer Mixes and KAPA Adapter plates are shipped with peelable seals. Replacement seals (three per plate) are peelable and pierceable.
* KAPA Universal Adapters, KAPA Universal UMI Adapters and KAPA UDI Adapters are not compatible with methyl-seq applications.