Qubit Value Lower Than UV Absorbance (Nucleic Acid Quantification)

Question

Accepted Answer

Qubit Fluorometer quantification value is significantly lower than UV absorbance (NanoDrop) measurement for the same sample Common causes: 1) Sample contaminated with molecules that absorb at 260 nm 2) Presence of degraded nucleic acids, small fragments, or free nucleotides 3) Protein contamination affecting UV absorbance reading 4) NanoDrop detecting contaminants while Qubit measures only intact nucleic acids 5) Poor sample mixing in assay tube Solutions: 1) Perform multiple Qubit assays (dsDNA, RNA HS, Protein) on same sample to determine composition 2) Check assay kit manual for list of potential interfering contaminants 3) Further dilute or purify sample to reduce contaminant concentration 4) Mix working solution and sample thoroughly in assay tube 5) Recognize that Qubit is more specific for intact nucleic acids than UV absorbance