Intracavity DNA Melting Analysis with Optofluidic Lasers

• Published in: Analytical chemistry (Washington) Vol. 84; no. 21; pp. 9558 - 9563
• Main Authors: Lee, Wonsuk, Fan, Xudong
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 06-11-2012
• Subjects: Analytical chemistry; Base Pair Mismatch; Chemistry; DNA - analysis; DNA - chemistry; DNA - genetics; Exact sciences and technology; Fluorescence; Genetic testing; Lasers; Microfluidic Analytical Techniques - methods; Nucleic Acid Denaturation; Optical Phenomena; Phase transitions; Signal transduction; Spectrometric and optical methods; Temperature effects; Transition Temperature; Temperature; Scanning; Melting; Nucleotide sequence; Device; Fluorescence; Melting curve; Phase transitions; Fluorescence spectrometry; Target; Signal; Dynamics; DNA; Quality; Laser; Lead; Detection; Differential method
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/ac302416g

DNA melting analysis holds great promise for simple and fast DNA sequence discrimination. However, conventional fluorescence-based methods suffer from a small differential signal and demanding melting curve analysis, both of which make it difficult to distinguish the target DNA from the mismatched one. Herein, we propose and demonstrate a highly specific intracavity DNA melting analysis scheme utilizing an optofluidic laser. The laser optically amplifies the small yet intrinsic thermal dynamic difference between the target and the single-base-mismatched DNA, resulting in a differential signal that is orders of magnitude greater than with fluorescence-based methods. In particular, the existence of a phase transition between the stimulated laser emission and fluorescence (i.e., spontaneous emission) enables accurate determination of the DNA transition temperature difference. Furthermore, the high differential signal in the intracavity detection allows for scanning of the laser excitation at a fixed temperature to distinguish two DNA sequences, which provides another means for rapid DNA analysis. In this paper, we first theoretically investigate DNA melting analysis using an optofluidic laser and then experimentally explore this scheme with a high-quality optofluidic ring resonator. Distinction of two DNA sequences of up to 100 bases long is demonstrated. The intracavity detection developed here will lead to novel optofluidic devices that enable rapid and simple analysis of DNAs with very long sequences.