In this collaboration, Meridian worked closely with the 4bases R&D team to optimize the NanoAdapter chemistry for lyophilization while maintaining its required performance within the Oxford Nanopore workflow.
APPLICATION NOTE
Integrated lyophilized NanoAdapter for simplified and robust library preparation workflows A collaboration between Meridian Life Science and 4bases SA R&D teams
Introduction 4bases is a biotechnology company specializing in the development of innovative library preparation kits and bioinformatics solutions for next-generation sequencing applications. Its products support multiple sequencing technologies and are designed to ensure compatibility with a wide range of sequencing platforms, including Oxford Nanopore Technologies.
Key benefits Lyophilized format Simplified handling
No cold-chain required Comparable sequencing performance
Figure 1. M1050 Lyophilized NanoAdapter Kit.
To further strengthen this compatibility and streamline the transition from a 4bases-prepared library to a sequencing- ready Oxford Nanopore library, 4bases developed and validated a dedicated final workflow step: the M1050 NanoAdapter Kit, designed for reliable and standardized use in clinical laboratory workflows. NanoAdapter is an integral part of the 4bases workflow. It performs end repair/A-tailing, adapter ligation and library cleanup, converting short-amplicon, long-amplicon and capture-based libraries into a format ready for the final Oxford Nanopore sequencing components. The prepared library can then use the Ligation Adapter and motor protein chemistry supplied in the Oxford Nanopore Technologies Ligation Sequencing Kit V14 (SQK-LSK114), enabling efficient nanopore capture and DNA translocation during sequencing. Meridian Life Science, the life science division of Meridian Bioscience, provides molecular biology reagents and assay development capabilities to diagnostic and life science manufacturers worldwide. Its advanced reagent stabilization platform combines formulation science with enzyme lyophilization expertise to support the development of ambient-stable molecular workflows.
Through this collaboration, Meridian worked closely with the 4bases R&D team to optimize the NanoAdapter chemistry for lyophilization while maintaining its required performance within the Oxford Nanopore workflow. The standard liquid M1050 NanoAdapter Kit had already been tested and validated across multiple 4bases applications. Building on this established performance, the Meridian Life Science and 4bases R&D teams jointly developed a lyophilized version of M1050, preserving its function within the validated workflow while introducing the practical advantages of a lyophilized reagent format. This Application Note evaluates the sequencing performance of the new lyophilized M1050 NanoAdapter Kit against the validated liquid formulation across representative 4bases library preparation workflows. Objective To confirm that the lyophilized M1050 NanoAdapter Kit maintains the validated performance of the liquid M1050 formulation and can be integrated as the final Oxford Nanopore compatibility step across different 4bases library preparation workflows.
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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
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APPLICATION NOTE
Results The M1050 Lyophilized NanoAdapter Kit demonstrated sequencing performance comparable to the validated liquid M1050 formulation across all evaluated 4bases library types. Representative results from each library type are shown in Figures 3–5. For both short- and long-amplicon libraries, sequencing yield, read count and fragment size distribution (N50) were highly consistent between the two NanoAdapter formulations. Similar pore occupancy profiles and Q score distributions further confirmed that the lyophilized format maintains compatibility with the Oxford Nanopore sequencing chemistry. Comparable results were also obtained for capture-based libraries, supporting the use of the lyophilized M1050 NanoAdapter Kit as a standardized final step for Oxford Nanopore compatibility across different 4bases library preparation strategies.
Figure 3. Representative sequencing results from a short-amplicon 4bases library processed with either the liquid M1050 NanoAdapter Kit or the M1050 Lyophilized NanoAdapter Kit.
Figure 4. Representative sequencing results from a long-amplicon 4bases library processed with either the liquid M1050 NanoAdapter Kit or the M1050 Lyophilized NanoAdapter Kit.
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APPLICATION NOTE
Across all tested applications, the lyophilized formulation maintained sequencing quality, pore utilization and overall workflow performance without requiring changes to the upstream 4bases library preparation protocols.
Figure 5. Representative sequencing results from a capture-based 4bases library processed with either the liquid M1050 NanoAdapter Kit or the M1050 Lyophilized NanoAdapter Kit.
Conclusion The M1050 NanoAdapter Kit connects 4bases library preparation workflows with Oxford Nanopore sequencing. By converting 4bases-prepared libraries into a format compatible with the Ligation Adapter and motor protein chemistry of the Oxford Nanopore Technologies Ligation Sequencing Kit V14 (SQK-LSK114), NanoAdapter enables a streamlined transition to nanopore sequencing. The liquid M1050 formulation had already been tested and validated across short- amplicon, long-amplicon and capture-based applications. The new lyophilized formulation, developed through the combined R&D expertise of Meridian Life Science and 4bases, generated comparable sequencing yield, read count, fragment size distribution, pore occupancy and sequencing quality. These results support the integration of the M1050 Lyophilized NanoAdapter Kit into existing 4bases workflows as a reliable, lyophilized solution that strengthens Oxford Nanopore compatibility while simplifying reagent handling, storage and transportation.
Key Benefits of the Lyophilized Formulation
Simplified reagent handling Increased reproducibility among experiments Improved storage and transportation
Compatible with existing workflows Validated across multiple applications Comparable sequencing performance
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