G4 Sequencing Platfom FAQs

Accurate quantification and proper quality check of NGS libraries are keys to successful sequencing runs. When quantifying libraries for sequencing on the Singular Genomics G4, we recommend using a fluorometric method such as Qubit or PicoGreen that utilize a double-strand DNA-specific fluorescent dye method to quantify the libraries. In addition, we recommend verifying average library size distribution through electrophoretic instrument analysis optimized for NGS (e.g. BioAnalyzer, or TapeStation). It is considered best practice to quantify individual libraries prior to pooling and loading in order to ensure the desired distribution of reads on the flow cell and optimum cluster density.

Library quantification using spectrophotometry-based methods should be avoided as any contaminating single-stranded nucleic acids and free nucleotides may lead to an overestimation of library concentration resulting in fewer clusters on the flow cell. Similarly, it is not recommended to solely use fragment analyzers (e.g. BioAnalyzer, TapeStation) to quantify Singular Genomics libraries as it may lead to an underestimation of library concentrations and over-seeded NGS runs.

See also the G4 Best Practices and Quality Control Guide for more information.

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