App Note - Integrated Lyophilized NanoAdapter_4Bases

APPLICATION NOTE

Study Design Short-amplicon, long-amplicon and capture-based libraries were generated using standard 4bases library preparation workflows. All upstream preparation steps and key enzymatic reactions were kept unchanged. Only the final NanoAdapter step was varied, using either the validated liquid M1050 formulation or the newly developed lyophilized M1050 formulation. (Fig. 2). The minor reconstitution adjustments required for the lyophilized format did not alter the overall workflow duration or the integration of NanoAdapter within the 4bases process.

Figure 2. Study design for comparing the liquid and lyophilized M1050 NanoAdapter formulations across amplicon- and capture- based 4bases workflows.

Following NanoAdapter processing, the libraries were completed using the relevant components of the Oxford Nanopore Technologies Ligation Sequencing Kit V14 (SQK- LSK114), sequenced on an Oxford Nanopore platform and compared using the following performance metrics:

Sequencing yield Read count N50

Q score distribution Pore occupancy over time The comparison included representative sequencing applications:

Short amplicon library Long amplicon library Capture-based library