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Specific DNA duplex formation at an artificial lipid bilayer: fluorescence microscopy after Sybr Green I staining

The article describes the immobilization of different probe oligonucleotides (4, 7, 10) carrying each a racemic mixture of 2,3-bis(hexadecyloxy)propan-1-ol (1a) at the 5’-terminus on a stable artificial lipid bilayer composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) and 1-palm...

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Autores principales: Werz, Emma, Rosemeyer, Helmut
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Beilstein-Institut 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4187062/
https://www.ncbi.nlm.nih.gov/pubmed/25298798
http://dx.doi.org/10.3762/bjoc.10.240
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author Werz, Emma
Rosemeyer, Helmut
author_facet Werz, Emma
Rosemeyer, Helmut
author_sort Werz, Emma
collection PubMed
description The article describes the immobilization of different probe oligonucleotides (4, 7, 10) carrying each a racemic mixture of 2,3-bis(hexadecyloxy)propan-1-ol (1a) at the 5’-terminus on a stable artificial lipid bilayer composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC). The bilayer separates two compartments (cis/trans channel) of an optical transparent microfluidic sample carrier with perfusion capabilities. Injection of unlabeled target DNA sequences (6, 8, or 9), differing in sequence and length, leads in the case of complementarity to the formation of stable DNA duplexes at the bilayer surface. This could be verified by Sybr Green I double strand staining, followed by incubation periods and thorough perfusions, and was visualized by single molecule fluorescence spectroscopy and microscopy. The different bilayer-immobilized complexes consisting of various DNA duplexes and the fluorescent dye were studied with respect to the kinetics of their formation as well as to their stability against perfusion.
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spelling pubmed-41870622014-10-08 Specific DNA duplex formation at an artificial lipid bilayer: fluorescence microscopy after Sybr Green I staining Werz, Emma Rosemeyer, Helmut Beilstein J Org Chem Full Research Paper The article describes the immobilization of different probe oligonucleotides (4, 7, 10) carrying each a racemic mixture of 2,3-bis(hexadecyloxy)propan-1-ol (1a) at the 5’-terminus on a stable artificial lipid bilayer composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC). The bilayer separates two compartments (cis/trans channel) of an optical transparent microfluidic sample carrier with perfusion capabilities. Injection of unlabeled target DNA sequences (6, 8, or 9), differing in sequence and length, leads in the case of complementarity to the formation of stable DNA duplexes at the bilayer surface. This could be verified by Sybr Green I double strand staining, followed by incubation periods and thorough perfusions, and was visualized by single molecule fluorescence spectroscopy and microscopy. The different bilayer-immobilized complexes consisting of various DNA duplexes and the fluorescent dye were studied with respect to the kinetics of their formation as well as to their stability against perfusion. Beilstein-Institut 2014-10-02 /pmc/articles/PMC4187062/ /pubmed/25298798 http://dx.doi.org/10.3762/bjoc.10.240 Text en Copyright © 2014, Werz and Rosemeyer https://creativecommons.org/licenses/by/2.0https://www.beilstein-journals.org/bjoc/termsThis is an Open Access article under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The license is subject to the Beilstein Journal of Organic Chemistry terms and conditions: (https://www.beilstein-journals.org/bjoc/terms)
spellingShingle Full Research Paper
Werz, Emma
Rosemeyer, Helmut
Specific DNA duplex formation at an artificial lipid bilayer: fluorescence microscopy after Sybr Green I staining
title Specific DNA duplex formation at an artificial lipid bilayer: fluorescence microscopy after Sybr Green I staining
title_full Specific DNA duplex formation at an artificial lipid bilayer: fluorescence microscopy after Sybr Green I staining
title_fullStr Specific DNA duplex formation at an artificial lipid bilayer: fluorescence microscopy after Sybr Green I staining
title_full_unstemmed Specific DNA duplex formation at an artificial lipid bilayer: fluorescence microscopy after Sybr Green I staining
title_short Specific DNA duplex formation at an artificial lipid bilayer: fluorescence microscopy after Sybr Green I staining
title_sort specific dna duplex formation at an artificial lipid bilayer: fluorescence microscopy after sybr green i staining
topic Full Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4187062/
https://www.ncbi.nlm.nih.gov/pubmed/25298798
http://dx.doi.org/10.3762/bjoc.10.240
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