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Nucleic Acid Quantification moderate

Negative Fluorescence Values

Symptom
Fluorescence reader displays negative values, which is physically impossible
Common Causes
  1. 1 Software autocorrecting for excessive background signal
  2. 2 Reader picking up and subtracting background light
  3. 3 Incompatible plate type with high autofluorescence
  4. 4 Excessive background interference
Solutions
  1. 1 Perform buffer-only control to assess background signal type
  2. 2 Switch to different plate type with lower autofluorescence
  3. 3 Use recommended tube types for optical clarity
  4. 4 Verify plate/tube compatibility with Qubit system
  5. 5 Check for sources of background light contamination
Related Video (2)
Bio-protocol Video ★ 85
Setting up the plate reader
"Directly demonstrates plate reader setup, which is essential for understanding how background corrections cause negative fluorescence values."
Bio-Rad Laboratories ★ 70
How to Use the New ChemiDoc MP for DNA Gel Imaging
"Shows imaging system software correction options, relevant to troubleshooting autocorrection artifacts in fluorescence measurements."
Source: thermofisher.com ↗
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