Determination of nucleic acids using fluorescence probe sensitized by microemulsion

Area of Science:

• Analytical Chemistry
• Biochemistry

Background:

• Nucleic acid determination is crucial in biological and chemical analyses.
• Existing fluorometric methods may lack sensitivity for trace-level detection.

Purpose of the Study:

• To establish a highly sensitive fluorometric method for quantifying nucleic acids.
• To investigate the interaction between azur A and nucleic acids.

Main Methods:

• Utilized the fluorescence quenching of azur A upon binding with nucleic acids.
• Employed Triton X-100 microemulsion to enhance assay sensitivity.
• Determined calibration curves and limits of detection for deoxyribonucleic acid (DNA).
• Studied the binding mode and calculated association constants.

Main Results:

• Achieved linear calibration curves for calf thymus DNA (0-6.0 microg/mL) and herring sperm DNA (0-7.0 microg/mL).
• Established high sensitivity with determination limits of 3.5 ng/mL for ct DNA and 3.8 ng/mL for hs DNA.
• Demonstrated successful application in analyzing nucleic acids in synthetic and real biological samples.

Conclusions:

• The developed azur A-based fluorometric method offers high sensitivity for nanogram-level nucleic acid determination.
• The use of Triton X-100 microemulsion significantly enhances detection sensitivity.
• The method is reliable for quantifying nucleic acids in diverse sample matrices.