Nucleic Acid Binding, Loading & Release
Circular Dichroism (CD) Spectroscopy
Conformational changes in DNA or siRNA upon binding to nanocarriers.
Purpose
To study conformational changes in nucleic acids (e.g., DNA double helix, siRNA) upon binding to nanocarriers.
Principle
Chiral molecules (like DNA/RNA) differentially absorb left- and right-circularly polarised light. Changes in the spectrum indicate structural perturbations (e.g., unwinding, condensation).
Standard protocol
- 1
Prepare samples
- Free nucleic acid.
- Complex at various N/P ratios.
- 2
Scan from 200–300 nm (DNA) or 200–260 nm (RNA).
- 3
Parameters: 1 nm bandwidth, 1 s response, 100 nm/min scan speed.
- 4
Subtract the buffer baseline.
Data interpretation
- B-DNA: +275 nm, −245 nm.
- Changes → condensation or structural distortion.
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