Library loading recommendations by assay for the AVITI24™ and the AVITI™ can be found in the Cloudbreak Sequencing User Guide.
AVITI24 delivers up to 50% increased sequencing output for 2x75 and 2x150 Cloudbreak™, Cloudbreak Freestyle™, and Trinity™ kits when running with AOS v3.4 or higher. Loading concentrations may need to be adjusted depending on the sequencing kit used and sequencing yield currently obtained on the AVITI.
For most 2x75 and 2x150 Cloudbreak and Cloudbreak Freestyle runs, we recommend a starting loading concentration of ~1.25X your current loading concentration. For example, if you typically load your library at 10pM to yield ~1 billion pass-filter reads with Cloudbreak Freestyle on AVITI, try starting with 12.5pM (a 25% increase) on AVITI24. To hit the 1.5B pass-filter polony metric, you should ideally target 1.9B raw polonies before filtering. Additional optimization of loading concentration may be required depending on your specific library type. If you have optimized your library output on Cloudbreak or Cloudbreak Freestyle runs on AVITI or AVITI24 software version < 3.4.0 to yield 1.1-1.2B pass-filter reads with standard run settings, you may not need to change your current loading concentration.
For Trinity kits, we recommend starting with your currently loading concentration without additional material.
Note that recommended loading concentrations are a starting point and can continue to be titrated upward as long as they maintain good quality and the output continues to increase. For more information on loading and quantification tips to help you maximize pass-filter reads from your sequencing runs, see What are some best practices for library quantification and loading to maximize my sequencing run density?
Additional information about increased sequencing output on AVITI24 can be found in the Knowledge Base article How does increased sequencing output on AVITI24 compare to performance on AVITI with and without Expert Mode HD? and the Technical Note Enhanced Output with AVITI24.