A Label-Free and Non-Destructive Separation Technique for Isolation of Targeted DNA from DNA Protein Mixture using Magnetic Au-Fe 3 O 4 Nanoprobes

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1 Supporting Information A Label-Free and Non-Destructive Separation Technique for Isolation of Targeted DNA from DNA Protein Mixture using Magnetic Au-Fe 3 O 4 Nanoprobes Ankan Dutta Chowdhury 1, 2, #,*, Nidhi Agnihotri 1, #, Ruey-an Doong 2, 3, Amitabha De 1 *E.mail: dc_ankan@yahoo.co.in, Tel: , Fax: Chemical Sciences Division, Saha Institute of Nuclear Physics, 1/AF Bidhannagar, Kolkata , India. 2 Institute of Environmental Engineering, National Chiao Tung University, 1001 University Road, Hsinchu 30010, Taiwan 3 Department of Biomedical Engineering and Environmental Sciences, National Tsing Hua University, 101 Section 2, Kuang-Fu Road, Hsinchu, 30013, Taiwan. Table of Contents: Table S1. Recent reports on Au-Fe 3 O 4 nanocomposites for targeting biomolecules. S2 Figure S1. Hydrodynamic diameter distribution of as-prepared Fe 3 O 4 and Au-Fe 3 O 4 nanocomposites by using dynamic light scattering measurement. S4 Figure S2. Melting temperature analysis of target DNA and duplex of probe ssdna by derivative of UV-visible curve. S5 Figure S3. UV-visible spectra of different concentration of target DNA in pure lysozyme protein mixture along with its corresponding separated DNA solutions after separation with nanoprobes. S6 S1

2 Table S1. Recent reports on Au-Fe 3 O 4 nanocomposites for targeting biomolecules: Name of published paper Main work frame References 1. Bifunctional Au-Fe 3 O 4 nanopartides for protein separation Protein separation, however no bioactivity was checked after separation ACS Nano, 2007, 1, Colorimetric aptasensing of ochratoxin A using Au@Fe 3 O 4 nanoparticles as signal indicator and magnetic separator cdna linkage for OTA sensing and separation Biosensors and Bioelectronics, 2016, 77, Three-Layer Magnetic Probes for DNA Composite Nanoparticle Journal of the American Chemical Society, 2005, 127, Ultrasensitive DNA monitoring by Au-Fe 3 O 4 nanocomplex Sensors and Actuators B: Chemical, 2012, 163, Manipulating the Power of an Additional Phase: A Flowerlike Au-Fe 3 O 4 Optical Nanosensor for Imaging Protease Expressions In vivo Oligomer conjugation for imaging ACS Nano, 2011, 5, Ultrasensitive electrochemical detection of DNA hybridization using Au/Fe 3 O 4 magnetic composites combined with silver enhancement Analyst, 2010, 135, Fe 3 O 4 /Au Core/Shell Nanoparticles Modified with Ni 2+ Nitrilotriacetic Acid Specific to Histidine-Tagged Proteins Protein separation, however no bioactivity was checked after separation Journal of Physical Chemistry C, 2010, 114, Dual-mode protein detection based on Fe 3 O 4 -Au hybrid nanoparticles Protein detection Nano Research, 2012, 5, S2

3 9. Fabrication and estimation of Au-coated Fe 3 O 4 nanocomposite powders for the separation and purification of biomolecules Glutathione sensing and separation Materials Science and Engineering: A, 2007, , Functionalization of magnetic gold/iron-oxide composite nanoparticles with oligonucleotides and magnetic separation of specific target and separation from water however no bioactivity was checked after separation Journal of Magnetism and Magnetic Materials, 2007, 311, Modification of DNA on magnetic beads DNA separation in water however no bioactivity was checked after separation US patent, US B2, Apr 19, Magnetic bead-based labelfree electrochemical detection of DNA hybridization Analyst, 2001, 126, A Label-Free and Non- Destructive Separation Technique for Isolation of Targeted DNA from DNA Protein Mixture using Magnetic Au-Fe 3 O 4 Nanoprobes in DNAprotein mixture (serum) and non-destructive separation, bioactivity of each component is crosschecked Current work S3

4 Intensity Fe 3 O 4 (372 nm) PDI: Au-Fe 3 O 4 (178 nm) PDI: Size (nm) Figure S1. Hydrodynamic diameter distribution of as-prepared Fe 3 O 4 and Au-Fe 3 O 4 nanocomposites by using dynamic light scattering measurement S4

5 da/dt target DNA ssdna-1+ssdna Temp( C) Figure S2. Melting temperature analysis of target DNA and duplex of probe ssdna by derivative of UV-visible curve. S5

6 Absorbance M DNA + Lysozyme Separated DNA 100 nm DNA + Lysozyme Separated DNA 10 nm DNA + Lysozyme Separated DNA Wavelength(nm) Figure S3. UV-Visible spectra of different concentration of target DNA in pure lysozyme protein mixture along with its corresponding separated DNA solutions after separation with nanoprobes. S6