Question: At what concentration should I load my libraries for Illumina sequencing?
Answer: Our recommendations for the optimal loading concentration, as well as pooling considerations, are listed in the sequencing section of each assay User Guide. Please ensure that the correct guide is used for the assay. Examples are shared below:
Single Cell ATAC and Single Cell Multiome ATAC & Gene Expression
Visium Spatial Gene Expression
Library concentration (nM or pM) should be determined by KAPA Library quantification kit (part no. KK4824) and size corrected using fragment analysis. For further details, please see: How do alternative quantification methods other than qPCR impact library loading?
While these loading concentrations have worked well in-house, there may be some optimization required for inherent differences between sequencing instruments. We recommend working with Illumina Technical Support to optimize cluster density.
Products: Single Cell Gene Expression, Single Cell Immune Profiling, Single Cell Multiome ATAC+GEX, Single Cell ATAC, Fixed RNA Profiling, Single Cell Gene Expression Flex, Visium for FFPE, Visium for Fresh Frozen, Visium CytAssist Spatial Gene Expression, Visium CytAssist Spatial Gene and Protein, Visium HD