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A solution for sample crosstalk and index hopping in multiplexed NGS

Unique, dual-matched adapters substantially reduce cross-talk errors

Webinar summary: Learn about the steps in NGS sample preparation that can lead to index hopping and sample crosstalk. Dr Kristina Giorda provides expert advice on how to reduce these artifacts at each point, providing cleaner, demultiplexed NGS data.

For most applications, the massive sequencing capabilities of current NGS platforms is far greater than the sequencing requirements for any single sample. Therefore, multiplexing several samples in a high-throughput sequencing run is an indispensable technique to maximize use of instrument capacity.

For multiplexing to be successful, artificial, sample-specific, index sequences are added during library preparation. After sequencing, during a process called demultiplexing, the indices are used to assign the millions of individual reads to the correct sample.

Historically, a relatively small number of unique sequences (8 x i5 and 12 x i7) have been used to create index combinations to identify multiplexed samples. Unfortunately, various mechanisms can cause the wrong index to be attached to some of the sequences from a sample. This physical addition of an index to the wrong sample is referred to as “index hopping”. With the combinatorial approach, a single index swap may cause a read to be mis-assigned to a different sample causing cross-talk.

In the webinar, Unique, dual-matched adapters mitigate index hopping between NGS samples, Dr Kristina Giorda explains the sources of sample cross-talk and discusses how unique, dual-matched adapters can facilitate the removal of errors from NGS data analysis. Throughout the presentation, Dr Giorda also offers advice for library preparation and target enrichment that can help reduce the chances of index hopping that contribute to sample cross-talk. Watch the webinar recording below to learn more about how to reduce sample cross-talk in your sequencing experiments.

For more information on these xGen® Dual Index UMI Adapters, or to place customized orders, please contact our scientific applications support group.

Published Aug 4, 2017