AVITI24 with OS v3.4.0 or later enhances sequencing performance and increases pass-filter reads by up to 50% for many sequencing applications. Depending on the application, some libraries benefit from a slight loading increase to reach this maximal output. Loading recommendations for different libraries are provided in the Cloudbreak Sequencing User Guide and additional information can be found in the Knowledge Base article How much library should I load with AVITI24 to maximize output?
For libraries that have previously been optimized for ~1 billion pass-filter reads using 2x150 and 2x75 Cloudbreak™ and Cloudbreak Freestyle™ kits, we recommend increasing loading concentration by ~25% as a starting point. Below we’ve provided an example comparison of a library sequenced with 1x loading on AVITI™, 1x loading on AVITI24, and 1.25x loading on AVITI24 to demonstrate the output enhancement with AVITI24 at 1x and 1.25x loading concentrations.
Comparison of Loading Concentrations: Whole-Exome Sequencing Libraries
Whole-exome sequencing (WES) libraries are a common PCR-plus library sequenced on AVITI. With AVITI24’s increased sequencing capacity, we assessed the impact of loading a WES library at the same amount on both AVITI and AVITI24, as well as with the increased (1.25x) loading concentration recommended for AVITI24 to maximize output.
With the same loading concentration (and same WES library), there is a 52% increase in pass-filter reads on AVITI24 (Figure 1A). A slightly higher loading concentration of 1.25x yields even more pass-filter reads while still maintaining high quality above 90% Q30 (Figure 1B).
For a comparison of AVITI24 with AVITI Expert Mode HD, see How does increased sequencing output on AVITI24 compare to performance on AVITI with and without Expert Mode HD? To see data from other assays sequenced with AVITI and AVITI24, see the Enhanced Output with AVITI24 Technical Note.