UV-Vis Spectrophotometry (Loading Efficiency & Quantification)
Nucleic acid concentration, loading capacity and encapsulation efficiency.
To quantify nucleic acid concentration and calculate loading efficiency, encapsulation efficiency and complexation yield.
Standard protocol
Method 1 — Direct measurement (if the carrier doesn’t interfere)
- 1
Disrupt the complex with 1% SDS or heparin.
- 2
Measure absorbance at 260 nm.
- 3
Calculate concentration using the extinction coefficient (e.g., dsDNA: 50 μg/mL = A260 of 1).
Method 2 — Dye-based assay (e.g., RiboGreen for RNA, PicoGreen for DNA)
- 1
Prepare a standard curve with free nucleic acid.
- 2
Treat samples with dye (protected from light).
- 3
Measure fluorescence (Ex/Em: 480/520 nm).
- 4
Back-calculate the amount of nucleic acid in the complex.
(Mass of loaded nucleic acid ÷ Mass of carrier) × 100%
(Actual loading ÷ Theoretical loading) × 100%
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