Why lyo-ready mixes are crucial for qPCR assay development

Molecular diagnostic laboratories often face significant and persistent issues throughout the quantitative PCR (qPCR) assay development process that can be resolved through lyophilization and lyo-ready mixes—simple solutions that effectively address these hurdles and move operations forward.

Current challenges in assay development

While traditional qPCR master mixes are formulated to provide sensitive and reproducible pathogen detection, they are not without limitations. For instance, master mix performance is reliant on optimal storage and transportation conditions, which can lead to complications when compromised. Poor reagent integrity can hinder assay performance and consequently delay validation studies, adding time to market entry. As such, it is important to incorporate methods and materials that reduce storage-related issues—such as lyophilization—into the development cycle.

Lyophilization is a common freeze-drying method that preserves a given reagent while maintaining its integrity. A lyophilization-ready, or lyo-ready, mix is a preformulated liquid mixture of enzyme(s) that can be easily combined with primers and probes and then lyophilized for on-demand use in a variety of applications—including qPCR. The right lyo-ready mix offers key advantages that help any lab overcome even the most pressing challenges when it meets industry standards.

Four key advantages when lyophilizing your qPCR reagents

The assay development process can be significantly enhanced by lyophilizing qPCR reagents, which provides several advantages:

• Long-term stability: Lyophilized reagents remain stable over extended periods of time without refrigeration, reducing risk of degradation and ensuring consistent, reliable performance once reconstituted.
• Easy transportation: Lyophilized mixes can withstand temperature fluctuations during shipping, which enhances their reliability, reduces costs, and ensures that they arrive in optimal condition for convenient, on-demand use.
• Cost-efficiency: By reducing the frequency of reagent preparation and product replacement, lyophilization significantly reduces material costs—making it easier to stay within budget while maintaining high-quality assay results.
• Sustainability: Because lyophilized qPCR mixes do not require refrigeration or ice packs during shipping and storage, they use less energy and packaging materials. This reduces their waste and energy consumption for more sustainable lab practices.

Top requirements for lyophilization

As with all laboratory techniques, lyophilization must adhere to industry best practices in order to be an effective tool. Optimal lyophilization meets the following requirements:

• Lyo-ready enzymes: Lyo-ready enzymes are specially formulated to withstand the lyophilization process and retain activity upon rehydration. This helps ensure that the lyophilized mixes perform correctly once reconstituted.
• Simplified preparation: A lab’s own primers and probes can be directly added to lyo-ready mixes without the need for other additives. This advantage streamlines the preparation process for a more efficient, less error-prone experience.
• High quality: Reproducible enzyme lots are essential for maintaining consistent and reliable assay results. Lyo-ready mixes should be manufactured in accordance with ISO 13485:2016 quality management standards in an FDA-registered facility. Some mixes are general purpose reagents (GPR) labeled "For Laboratory Use," providing an added level of confidence in laboratory results.
• Performance in multiplex assays: Certain lyo-ready mixes can be used in multiplex assays (Fig. 1), which ultimately allows users to process more samples in less time.

Get started with a free sample

Takara Bio offers a variety of mixes in lyo-ready and lyo-cake formats that are ideally formulated for your qPCR processes.

Request your free evaluation samples today.