TY - JOUR
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 - http://www.scopus.com/inward/record.url?scp=77049099770&partnerID=8YFLogxK
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 -
