How much library should be loaded for G4 runs?

This article summarizes the loading concentration recommendations for the G4 Sequencing Platform

The optimal loading concentration will depend on the library type and on the Sequencing kit used (F2 or F3), and we recommend a titration run to determine empirically the optimal loading concentration per library type.

Please use our G4 Quick Reference Card to help guide your titration experiment. In general, we see loading concentrations between 10-30pM, loading 260uL per lane. Please note that the relationship between cluster density and loading concentration is nonlinear, so small, empirical adjustments are necessary to determine the optimum loading concentration.

Please use our G4 Loading Concentration Calculator to help you set up your dilutions.

The titration experiment can be done across one flow cell. As each Sample Cartridge well corresponds to a specific lane on the flow cell, it is important that the sample location matches the sample data sheet. Use our QRC to select 4 of the “Loading Concentrations” you would like to test. After the run completes, look at the results to see which concentration gives the optimal output and sequencing metrics.

Please contact Customer Care if you would like to discuss loading concentration optimization.