Temperature-Related Reading Errors (Nucleic Acid Quantification)

Question

Accepted Answer

Inconsistent or inaccurate readings caused by temperature variations in samples, standards, or reagents Common causes: 1) Buffer and Qubit reagent not at room temperature (stored at 4°C) 2) Samples too warm from centrifuge 3) Tubes warming up inside instrument during extended readings 4) Holding tubes in hand too long before reading 5) Temperature fluctuations between samples and standards 6) Assay is temperature sensitive - fluorescent signal decreases at higher temperatures Solutions: 1) Store buffer and Qubit reagent in DMSO at room temperature, not refrigerated 2) Allow buffer previously stored at 4°C to warm for 2-3+ hours to reach room temperature 3) Let samples from centrifuge cool to room temperature before measurement 4) Remove tubes from instrument between multiple readings 5) Allow tubes to equilibrate to room temperature for 30 seconds before rereading 6) Do not hold tubes in hand for extended time before reading 7) Perform entire assay at consistent room temperature 8) Ensure samples and working solution are not too warm