Standards Incorrect Message on Qubit Fluorometer (Nucleic Acid Quantification)

Question

Accepted Answer

Qubit Fluorometer displays 『Standards Incorrect』 error message during calibration or measurement Common causes: 1) Standard 2 fluorescence value not 10-50 fold higher than Standard 1 2) For protein assay, Standard 3 value not approximately twice Standard 2 3) Standards used in incorrect order 4) Incorrect volumes used: should be 10 µL standard + 190 µL working solution 5) RNA standard degraded in opened tube 6) Qubit assay kit older than one year or expired 7) Instrument malfunction - red/blue light not flashing 5 times during calibration Solutions: 1) Check raw fluorescence values under 『Check Standards』 or 『Check Calibration』 2) Verify standards used in correct order 3) Confirm 10 µL standard + 190 µL working solution volumes 4) For RNA assays, use unopened Standard 2 tube 5) Replace kit if older than 6 months from receipt or over one year 6) Test instrument by lifting lid and running new calibration - verify light flashes 5 times