DNA-magnetic Particle Binding Analysis by Dynamic and Electrophoretic Light Scattering

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F62156489%3A43210%2F17%3A43912860" target="_blank" >RIV/62156489:43210/17:43912860 - isvavai.cz</a>

Alternative codes found

RIV/00216305:26620/17:PU126240

Result on the web

<a href="https://doi.org/10.3791/56815" target="_blank" >https://doi.org/10.3791/56815</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.3791/56815" target="_blank" >10.3791/56815</a>

Result language

angličtina

Original language name

DNA-magnetic Particle Binding Analysis by Dynamic and Electrophoretic Light Scattering

Original language description

Isolation of DNA using magnetic particles is a field of high importance in biotechnology and molecular biology research. This protocol describes the evaluation of DNA-magnetic particles binding via dynamic light scattering (DLS) and electrophoretic light scattering (ELS). Analysis by DLS provides valuable information on the physicochemical properties of particles including particle size, polydispersity, and zeta potential. The latter describes the surface charge of the particle which plays major role in electrostatic binding of materials such as DNA. Here, a comparative analysis exploits three chemical modifications of nanoparticles and microparticles and their effects on DNA binding and elution. Chemical modifications by branched polyethylenimine, tetraethyl orthosilicate and (3-aminopropyl) triethoxysilane are investigated. Since DNA exhibits a negative charge, it is expected that zeta potential of particle surface will decrease upon binding of DNA. Forming of clusters should also affect particle size. In order to investigate the efficiency of these particles in isolation and elution of DNA, the particles are mixed with DNA in low pH (similar to 6), high ionic strength and dehydration environment. Particles are washed on magnet and then DNA is eluted by Tris-HCl buffer (pH = 8). DNA copy number is estimated using quantitative polymerase chain reaction (PCR). Zeta potential, particle size, polydispersity and quantitative PCR data are evaluated and compared. DLS is an insightful and supporting method of analysis that adds a new perspective to the process of screening of particles for DNA isolation.

Czech name

—

Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

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OECD FORD branch

10609 - Biochemical research methods

Project

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year

2017

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Journal of Visualized Experiments

ISSN

1940-087X

e-ISSN

—

Volume of the periodical

Neuveden

Issue of the periodical within the volume

129

Country of publishing house

US - UNITED STATES

Number of pages

9

Pages from-to

&quot;nestrankovano&quot;

UT code for WoS article

000417688700101

EID of the result in the Scopus database

2-s2.0-85034053223