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Monika Hassel Michael G. Cornelius Jochen vom Brocke Heinz H. Schmeiser 《Electrophoresis》2010,31(2):299-302
The model organism Hydra has been used for molecular studies for more than 20 years, however, its DNA base composition has not been determined yet. We have analyzed DNA and total RNA of the freshwater polyp Hydra magnipapillata with two independent procedures of high accuracy and sensitivity – fluorescence labeling of nucleotides followed by CE‐LIF detection and 32P‐postlabeling. DNA of Hydra was digested either to deoxyribonucleoside‐5′‐monophosphates or deoxyribonucleoside‐3′‐monophosphates selectively conjugated with the fluorescent dye 4,4‐difluoro‐5,7‐dimethyl‐4‐bora‐3a,4a‐diaza‐s‐indacene‐3‐propionyl ethylene diamine hydrochloride (BODIPY FL EDA) separated and detected using CE‐LIF. Both versions of the assay revealed a high A+T composition of 78 and 71%, whereas total DNA methylation (5‐methyldeoxycytidine) was 2.6 and 3.1%. Total Hydra RNA showed highest base levels for guanine (33%) and a level of 1.4% for pseudouracil. All values were in good agreement with those determined by the 32P‐postlabeling method. 相似文献
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We introduce a new metric in the space of fuzzy polynucleotides. 相似文献
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H.-K. Kim E. Tuite B. Nordén B.W. Ninham 《The European physical journal. E, Soft matter》2001,4(4):411-417
The source of the activation energy that allows cutting of DNA by restriction enzymes is unclear. A systematic study of the
cutting efficiency of the type-II restriction endonuclease EcoRI, with varying background electrolyte ion pair and buffer reported here, shows only a modest dependence of efficiency on
cation type. Surprisingly, efficiency does depend strongly on the presumed indifferent anion of the background salt. What
emerges is that competition between the background salt anion and the buffer anion for the enzyme and DNA surfaces is crucial.
The results are unexpected and counterintuitive from the point of view of conventional electrolyte theory. However, taken
together with recent developments in surface chemistry, the results do fall into place and could also suggest a novel mechanism
for enzyme activity as an alternative to metal-activated hydrolysis: microscopic cavitation in a hydrophobic pocket might
be the source of activation energy.
Received 19 June 2000 and Received in final form 17 October 2000 相似文献
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