Exonuclease III action on microarrays: Observation of DNA degradation by fluorescence correlation spectroscopy

Nicky Ehrlich; Katrin Anhalt; Christian Hübner; Susanne Brakmann

doi:10.1016/j.ab.2009.12.004

Exonuclease III action on microarrays: Observation of DNA degradation by fluorescence correlation spectroscopy

Nicky Ehrlich, Katrin Anhalt, Christian Hübner, Susanne Brakmann^*

^*Korrespondierende/r Autor/-in für diese Arbeit

Institut für Physik

7 Zitate (Scopus)

Abstract

DNA with all cytosines, thymines, or all pyrimidines of one strand substituted by fluorescently labeled analogs shows diminished solubility in aqueous media and a strong tendency to aggregation that hampers enzymatic downstream processing. In this study, immobilization of fully fluorescently labeled DNA on microarrays was shown to resolve the named problems and to enable successive DNA degradation by exonuclease III. Fluorescence correlation spectroscopy and single-molecule counting for monitoring the course of DNA hydrolysis in real time revealed the virtually processive degradation of labeled DNA that occurred at an average rate of approximately 4 nt/s.

Originalsprache	Englisch
Zeitschrift	Analytical Biochemistry
Jahrgang	399
Ausgabenummer	2
Seiten (von - bis)	251-256
Seitenumfang	6
ISSN	0003-2697
DOIs	https://doi.org/10.1016/j.ab.2009.12.004
Publikationsstatus	Veröffentlicht - 15.04.2010

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10.1016/j.ab.2009.12.004

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title = "Exonuclease III action on microarrays: Observation of DNA degradation by fluorescence correlation spectroscopy",

abstract = "DNA with all cytosines, thymines, or all pyrimidines of one strand substituted by fluorescently labeled analogs shows diminished solubility in aqueous media and a strong tendency to aggregation that hampers enzymatic downstream processing. In this study, immobilization of fully fluorescently labeled DNA on microarrays was shown to resolve the named problems and to enable successive DNA degradation by exonuclease III. Fluorescence correlation spectroscopy and single-molecule counting for monitoring the course of DNA hydrolysis in real time revealed the virtually processive degradation of labeled DNA that occurred at an average rate of approximately 4 nt/s.",

author = "Nicky Ehrlich and Katrin Anhalt and Christian H{\"u}bner and Susanne Brakmann",

year = "2010",

month = apr,

day = "15",

doi = "10.1016/j.ab.2009.12.004",

language = "English",

volume = "399",

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journal = "Analytical Biochemistry",

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T1 - Exonuclease III action on microarrays

T2 - Observation of DNA degradation by fluorescence correlation spectroscopy

AU - Ehrlich, Nicky

AU - Anhalt, Katrin

AU - Hübner, Christian

AU - Brakmann, Susanne

PY - 2010/4/15

Y1 - 2010/4/15

N2 - DNA with all cytosines, thymines, or all pyrimidines of one strand substituted by fluorescently labeled analogs shows diminished solubility in aqueous media and a strong tendency to aggregation that hampers enzymatic downstream processing. In this study, immobilization of fully fluorescently labeled DNA on microarrays was shown to resolve the named problems and to enable successive DNA degradation by exonuclease III. Fluorescence correlation spectroscopy and single-molecule counting for monitoring the course of DNA hydrolysis in real time revealed the virtually processive degradation of labeled DNA that occurred at an average rate of approximately 4 nt/s.

AB - DNA with all cytosines, thymines, or all pyrimidines of one strand substituted by fluorescently labeled analogs shows diminished solubility in aqueous media and a strong tendency to aggregation that hampers enzymatic downstream processing. In this study, immobilization of fully fluorescently labeled DNA on microarrays was shown to resolve the named problems and to enable successive DNA degradation by exonuclease III. Fluorescence correlation spectroscopy and single-molecule counting for monitoring the course of DNA hydrolysis in real time revealed the virtually processive degradation of labeled DNA that occurred at an average rate of approximately 4 nt/s.

UR - https://www.scopus.com/pages/publications/77049099770

U2 - 10.1016/j.ab.2009.12.004

DO - 10.1016/j.ab.2009.12.004

M3 - Journal articles

C2 - 20004637

AN - SCOPUS:77049099770

SN - 0003-2697

VL - 399

SP - 251

EP - 256

JO - Analytical Biochemistry

JF - Analytical Biochemistry

IS - 2

ER -

Exonuclease III action on microarrays: Observation of DNA degradation by fluorescence correlation spectroscopy

Abstract

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