Check your RNA quality with excellent speed and accuracy on Lunatic

Significance of the Topic

The high purity of RNA is crucial for downstream applications such as RNA-LNP therapeutics, qRT-PCR, and RNA-seq, where DNA and protein contaminants can compromise specificity and sensitivity.

Objectives and Study Overview

This application note demonstrates Lunatic, a next-generation UV/Vis spectrometer optimized for rapid and accurate RNA quantification and purity assessment, including deconvolution of contaminant signals from DNA, proteins, and extraction reagents.

Methodology and Used Instrumentation

• RNA and DNA samples: Calf liver RNA purified and calf thymus DNA diluted to target concentrations
• Contamination studies: RNA spiked with varying DNA ratios (5–40%), BSA protein (10–50% v/v) and serial dilutions
• Measurements: Lunatic UV/Vis system with High Lunatic plates, 2 µL samples, up to 96 per run
• Data analysis: Unmix algorithm to separate absorbance contributions of RNA, DNA, protein, and turbidity; regression and IQR outlier removal performed in RStudio/R

Main Results and Discussion

• Precision: Six replicates of 1,000 ng/µL pure RNA showed low CV and SD across absorbance spectra.
• DNA contamination: Unmix accurately quantifies RNA down to 60% purity, with R2 = 0.9925 for measured vs. target RNA in mixed samples.
• Concentration range: Reliable RNA percentage (70±2%) across total nucleic acid concentrations from 62.5 to 1,000 ng/µL.
• Protein interference: BSA-spiked samples produced correct RNA concentrations reflecting dilution by protein, demonstrating robust deconvolution.

Benefits and Practical Applications

The Lunatic platform streamlines RNA quality control by eliminating dyes and standard curves, reducing pipetting errors, and delivering high-throughput measurements in under 10 minutes. This enhances workflow efficiency in academic and industrial settings, ensuring accurate nucleic acid quantification for genomics, therapeutics, and QA/QC laboratories.

Future Trends and Opportunities

• Integration with automated liquid handlers for end-to-end high-throughput sample processing.
• Expansion of Unmix algorithms to additional biomolecules and complex matrices.
• Adoption in clinical environments for rapid RNA-based diagnostic assays.
• Potential development of cloud-based data management and quality monitoring dashboards.

Conclusion

Lunatic UV/Vis spectrophotometry with Unmix technology provides a robust, precise, and user-friendly solution for RNA quantification and purity assessment, reliably detecting DNA and protein contaminants across broad concentration ranges without additional reagents or complex protocols.

Used Instrumentation

• Unchained Labs Lunatic UV/Vis system with High Lunatic plates
• 100 kDa centrifugal filter (Merck)

References

• Chomczynski, P. & Sacchi, N. Anal. Biochem. 162, 156–159 (1987).