Abstract
| Original language | English |
|---|---|
| Title of host publication | Nucleic Acid Detection and Structural Investigations : Methods and Protocols |
| Editors | Kira Astakhova , Syeda Atia Bukhari |
| Number of pages | 8 |
| Volume | 2063 |
| Publisher | Springer |
| Publication date | 2020 |
| Pages | 37-44 |
| Chapter | 4 |
| ISBN (Print) | 978-1-0716-0137-2 |
| ISBN (Electronic) | 978-1-0716-0138-9 |
| DOIs | |
| Publication status | Published - 2020 |
| Series | Methods in Molecular Biology |
|---|---|
| ISSN | 1064-3745 |
Keywords
- DNA oncogene
- Fluorescent oligonucleotides
- Hybridization assay
- Fluorometry
Cite this
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Solid-Phase Hybridization Assay for Detection of Mutated Cancer DNA by Fluorescence. / Taskova, Maria; Astakhova, Kira.
Nucleic Acid Detection and Structural Investigations: Methods and Protocols. ed. / Kira Astakhova; Syeda Atia Bukhari. Vol. 2063 Springer, 2020. p. 37-44 (Methods in Molecular Biology).Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review
TY - CHAP
T1 - Solid-Phase Hybridization Assay for Detection of Mutated Cancer DNA by Fluorescence
AU - Taskova, Maria
AU - Astakhova, Kira
PY - 2020
Y1 - 2020
N2 - 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.
AB - 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.
KW - DNA oncogene
KW - Fluorescent oligonucleotides
KW - Hybridization assay
KW - Fluorometry
U2 - 10.1007/978-1-0716-0138-9_4
DO - 10.1007/978-1-0716-0138-9_4
M3 - Book chapter
SN - 978-1-0716-0137-2
VL - 2063
SP - 37
EP - 44
BT - Nucleic Acid Detection and Structural Investigations
A2 - , Kira Astakhova
A2 - , Syeda Atia Bukhari
PB - Springer
ER -