Enhanced Chemiluminescent Detection of Fluorescein-Labeled Nucleic Acids Compared to ³²P Labeling Methods: Lactoferrin as a Marker

Abstract

This study evaluated the ability of nonradioactive, sensitive Northern blotting methods to detect small amounts of messenger RNA. The more traditional ³²P labeling and detection method was compared to enhanced chemiluminescent (ECL) detection of fluorescein labeled nucleic acids. The results show that the ECL method is equally as sensitive as ³²P, and both methods are highly quantitative by image analysis. The fluorescein generated probes can be stored for several months as compared to several days storage for ³²P labeled probes. The ECL method of detecting fluorescein labeled nucleic acid probes should prove useful for many applications in biochemistry, molecular biology, and toxicology.

Key Words: :

• Northern blotting
• Enhanced chemiluminescence (ECL)
• Fluorescein random prime labeling
• Lactoferrin (LF)
• Phosphorous-32 (³²P)