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

Yazan Haddad; Simona Dostalova; Jiri Kudr; Ondrej Zitka; Zbynek Heger; Vojtech Adam

doi:10.3791/56815

Journal

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Chemistry

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

JoVE Journal
Chemistry

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JoVE Journal Chemistry

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

11,960 Views

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10:35 min

•

November 09, 2017

DOI:

10.3791/56815-v

DOI:

10.3791/56815-v

•

10:35 min

November 09, 2017

•

1 Views

Yazan Haddad^1,2, Simona Dostalova^1,2, Jiri Kudr^1,2, Ondrej Zitka^1,2, Zbynek Heger^1,2, Vojtech Adam^1,2

¹Department of Chemistry and Biochemistry,Mendel University in Brno, ²Central European Institute of Technology,Brno University of Technology

Transcript

The overall goal of this experiment is to analyze binding of DNA and Magnetic Particles via Dynamic and Electrophoretic Light Scattering.This method can help answer key questions in the Biotechnology field such as, how fast and strong is the binding between nucleic acid and magnetic particles.The main advantage of this technique is the visualization of physical-chemical changes on the particles such as size, polydispersity or zeta potential.The implications of this technique extend towards molecular diagnosis because of increasing demand for fast and sensitive DNA isolation.In this procedure, add 20 millimoles of Ferric Chloride and ten millimoles of Ferrous Chloride in 20 milliliters of Deoxygenated Double Distilled Water.Next, stir the obtained solution vigorously in beaker under Nitrogen gas atmosphere using a mechanical stirrer until a transparent solution is obtained.Then, under rapid mechanical stirring, add 150 milliliters of one molar of aqueous ammonia drop by drop using dropping funnel over an hour.Afterward, transfer the solution into a screwable vessel and heat the solution at 75

Summary

This protocol describes the synthesis of magnetic particles and evaluation of their DNA-binding properties via dynamic and electrophoretic light scattering. This method focuses on monitoring changes in particle size, their polydispersity and zeta potential of particle surface which play major role in binding of materials such as DNA.