Solid-Phase Hybridization Assay for Detection of Mutated Cancer DNA by Fluorescence

Maria Taskova, Kira Astakhova*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingBook chapterResearchpeer-review

Abstract

We report a straightforward protocol for the detection of mutated DNA extracted from cancer cells. The assay combines a step-wise solid-phase hybridization and a readout by fluorescence emission. We detect a single-nucleotide polymorphism in two human oncogenes, BRAF and EGFR, and reach a limit of the detection of 300 pM by conventional fluorometry. The protocol described herein may be used as a foundation for development of automatic optimized assays capable for detection of mutant DNA and RNA in vitro and in cells.
Original languageEnglish
Title of host publicationNucleic Acid Detection and Structural Investigations : Methods and Protocols
EditorsKira Astakhova , Syeda Atia Bukhari
Number of pages8
Volume2063
PublisherSpringer
Publication date2020
Pages37-44
Chapter4
ISBN (Print)978-1-0716-0137-2
ISBN (Electronic)978-1-0716-0138-9
DOIs
Publication statusPublished - 2020
SeriesMethods in Molecular Biology
ISSN1064-3745

Keywords

  • DNA oncogene
  • Fluorescent oligonucleotides
  • Hybridization assay
  • Fluorometry

Cite this

Taskova, M., & Astakhova, K. (2020). Solid-Phase Hybridization Assay for Detection of Mutated Cancer DNA by Fluorescence. In K. A., & S. A. B. (Eds.), Nucleic Acid Detection and Structural Investigations: Methods and Protocols (Vol. 2063, pp. 37-44). Springer. Methods in Molecular Biology https://doi.org/10.1007/978-1-0716-0138-9_4