Fiber optic system for rapid analysis of amplified DNA fragments

J. Matthew Mauro; Lynn Kim Cao; Joel P. Golden

doi:10.1117/12.237615

1 April 1996 Fiber optic system for rapid analysis of amplified DNA fragments

J. Matthew Mauro, Lynn Kim Cao, Joel P. Golden

Proceedings Volume 2680, Ultrasensitive Biochemical Diagnostics; (1996) https://doi.org/10.1117/12.237615
Event: Photonics West '96, 1996, San Jose, CA, United States

Abstract

We have developed a fiber optic sensor for rapid and direct analysis of PCR-amplified DNA fragments with minimal sample processing and real-time data readout. To accomplish this, a novel DNA-recognition system was built onto the surface of fused silica fibers. DNA fragments, labeled with a fluorophore during amplification, are bound to and detected at the fiber surface by means of evanescent wave excitation/emission. Excess unincorporated fluorescent single-stranded oligonucleotide PCR primers make only a small contribution to the signal, as the modified fiber surface only efficiently binds double-stranded DNA with the proper PCR-incorporated terminal nucleotide sequence (5'-ATGACTCAT-3'). The surface- bound double-stranded DNA recognition element utilizes a genetically engineered dimeric sequence-specific DNA binding protein. Self-assembly into the proper conformation for binding DNA occurs by means of specific interactions of the active dimer with the F_c domains of a layer of IgG molecules (antibodies) covalently attached directly to the fiber surface. The modified fiber surface is regenerated between samples by stripping away bound DNA with high salt concentrations.

Citation Download Citation

J. Matthew Mauro, Lynn Kim Cao, and Joel P. Golden "Fiber optic system for rapid analysis of amplified DNA fragments", Proc. SPIE 2680, Ultrasensitive Biochemical Diagnostics, (1 April 1996); https://doi.org/10.1117/12.237615

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