Best Practice for Nucleic Acid Thermal Stability Measurements Using the Cary 3500 UV-Vis Spectrophotometer - Thermal melt (Tm) analysis using rapid, precise temperature-dependent UV-Vis absorbance measurements
UV-Vis spectrophotometers have been used widely for nucleic acid quantification and quality control (QC) utilizing the fact that nucleic acids have a maximum absorbance at 260 nm (1). The concentration of nucleic acids can be easily estimated using the absorbance at 260 nm and the established absorption coefficient. Often a background correction is also performed, for example collecting a baseline using a solution containing everything but the nucleic acid or by measuring the absorbance at a wavelength that nucleic acids do not absorb. Double stranded nucleic acids are bound by hydrogen bonds between the base pairs. The temperature at which double stranded nucleic acids denature to become single stranded depends on the: – sequence and length of the nucleic acid – the pH and buffer conditions – and any mismatches in base pairs between the two strands As such, the melting temperature is very useful analytical tool and can be studied by monitoring the absorbance at 260 nm as temperature is increased or decreased. As the temperature is increased, the hydrogen bonds between the strands are broken and the double stranded nucleic acid separates into two
GMP Validated Next Generation Sequencing Services: Viral Safety Testing & Genetic Characterization
March 26th 2025This brochure will expand on our NGS testing services to biopharmaceutical and biotech companies, including viral contamination testing and genetic characterization of cell lines, bulk harvests, virus seeds, and raw materials used for production of biologics including cell and gene therapy products.
Develop Potency Assays Early for ATMP Success
March 26th 2025Early potency assay development and MOA understanding are critical for ATMP success. Delays can increase costs and risk failure, but recent advances highlight the need for timely, validated approaches to ensure regulatory and commercial success.
