Supplementary Data
S1.: Expression of procedures of paramagnetic microparticles crafting
Paramagnetic microparticles MAN16 were crafted as it follows: 10 g of FeCl3·6H2O was
dissolved in 800 mL of water (m/v). Subsequently 2 g of NaBH4 in 100 mL of 3.5% NH3
(m/v) was added. Mixture was heated at 100 ºC for 2 hours and after cooling the external
magnet (Chemagen, Baesweiler, Germany) was used for washing with water. Final volume of
solution was 100 mL. 20 mL of crafted solution was mixed with 1 g of Dowex 50WX4-400
(m/v) (Sigma-Aldrich, St. Louis, MO, USA). Resulting product was shaken on Biosan OS-10
(Biosan, Riga, Latvia) overnight. Finally, the product was separated using external magnetic
field, washed with water and dried at 40 ºC.
Paramagnetic microparticles MAN18 were crafted as it follows: 10 g of FeCl3·6H2O was
dissolved in 800 mL of water (m/v). Subsequently 2 g of NaBH4 in 100 mL of 3.5% NH3
(m/v) was added. Mixture was heated at 100 ºC for 2 hours and after cooling the external
magnet (Chemagen, Baesweiler, Germany) was used for washing with water. Final volume of
solution was 100 mL. 20 mL of crafted solution was mixed with 2 mL of titanium
isopropoxide (v/v) (Sigma-Aldrich, St. Louis, MO, USA) in concentration of 1 mg.mL-1.
Resulting product was shaken on Biosan OS-10 (Biosan, Riga, Latvia) overnight. Finally, the
product was separated using external magnetic field, washed with water and dried at 40 ºC.
Paramagnetic microparticles MAN38 were crafted as it follows: 7.5 g of FeNO3·9H2O was
dissolved in 400 mL of water (m/v). Subsequently 1 g of NaBH4 in 50 mL of 3.5% NH3 (m/v)
was added. Mixture was heated at 100 ºC for 2 hours and after cooling the external magnet
(Chemagen, Baesweiler, Germany) was used for washing with water. Final volume of
solution was 100 mL. 75 mL of solution was mixed with 25 mL of isopropanol (v/v) (SigmaAldrich, St. Louis, MO, USA). Mixture was separated using external magnetic field and then
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150 mL of isopropanol was added again (m/v). Subsequently was amonnia (20 mL, 28%)
added (v/v) and slowly 3.4 mL of 3-aminopropyltriethoxysilane (Sigma-Aldrich, St. Louis,
MO, USA) was added (v/v). Mixture was shaken at 40 ºC for 2 hours and then at 25 ºC
overnight. The precipitate was separated using external magnetic field and washed 2 times
with 75 mL of ethanol. Finally, the product was dried at 40 ºC.
Paramagnetic microparticles MAN51 were crafted as it follows: 7.5 g of FeNO3·9H2O was
dissolved in 400 mL of water (m/v). Subsequently 1 g of NaBH4 in 50 mL of 3.5% NH3 (m/v)
was added. Mixture was heated at 100 ºC for 2 hours and after cooling the external magnet
(Chemagen, Baesweiler, Germany) was used for washing with water. Final volume of
solution was 100 mL. 20 mL of crafted solution was mixed with 1 g of amberlite XAD4 20 60 mesh (m/v) (Sigma-Aldrich, St. Louis, MO, USA). Resulting product was shaken on
Biosan OS-10 (Biosan, Riga, Latvia) overnight. Finally, the product was separated using
external magnetic field, washed with water and dried at 40 ºC.
Paramagnetic microparticles MAN59 were crafted as it follows: 7.5 g of FeNO3·9H2O was
dissolved in 400 mL of water (m/v). Subsequently 1 g of NaBH4 in 50 mL of 3.5% NH3 (m/v)
was added. Mixture was heated at 100 ºC for 2 hours and after cooling the external magnet
(Chemagen, Baesweiler, Germany) was used for washing with water. Final volume of
solution was 100 mL. 20 mL of solution was mixed with 2 mL of 18% poly(4-styrenesulfonic
acid) (v/v) (Sigma-Aldrich, St. Louis, MO, USA). Mixture was shaken for 8 hours and 3 times
washed with water using external magnetic field. Finally, the mixture was dried at 40 ºC.
S2.: Escherichia coli transformation and GFP production
For Escherichia coli transformation, the pGLO plasmid (Bio-Rad, Philadelphia, PA, USA),
containing the reporter gene for GFP obtained from jellyfish Aequorea victoria, was used.
GFP gene was under control of the arabinose-induced promoter araBAD and the araC gene
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encoding regulator protein. The chemical transformation was carried out using Invitrogen
Competent according to the manufacturer’s protocol and TOP10 Escherichia coli was used as
a host strain (Life Technologies, Carlsbad, CA, USA). Bacteria transformed with pGLO
plasmid were selected according to their ampicillin resistance.
The positive transformants were grown in Luria-Bertani broth with 100 mg.L-1 of ampicillin
and 0.2% arabinose (w/w) and incubated at 32 °C overnight. Bright green cells were harvested
using centrifuge type 5340R (Eppendorf AG, Hamburg, Germany) at 4000 rpm for 10 min.
The pellet was resuspended thoroughly by rapidly pipetting using TE buffer (1 mM EDTA
with pH 8, 10 mM TRIS-HCl pH 7.5). To initiate enzymatic digestion of the bacterial cell
wall 10 mg.mL-1 of fresh lysozyme (Sigma Aldrich) was added and then the resuspended
bacterial pellet was frozen. The protein fraction was harvested by centrifugation for 10
minutes at 14000 rpm and 4 °C.
GFP was further purified from the bacterial lysate using fast protein liquid chromatography
system Biologic DuoFlow (Bio-Rad, Philadelphia, PA, USA) consisting of two
chromatographic pumps for transport of elution buffers, gel filtration column (Macro-Prep
Methyl HIC support, Bio-Rad, Philadelphia, PA, USA), injection valve with 1 mL injection
loop, UV-VIS detector and fraction collector (Bio-Rad, Philadelphia, PA, USA). TE buffer
(pH 8) was used as a mobile phase at a flow rate of 6 mL.min-1. Separation of GFP was done
using gradient elution method based on gradual decrease of ammonium sulphate
concentration. UV detection was carried out at 280 nm. Fractions were collected
approximately in volume of 1 mL. Total analysis time was 30 minutes.
S3.: Procedure of preparation of magnetic dsDNA(Th) complex
10 µL of Dynabeads Oligo (dT)25 (Invitrogen, Oslo) was dosed into each well of the PCR
plate (PCR 96, Eppendorf, Hamburg, Germany). Then, the beads were subsequently washed
three times with 100 µL of phosphate buffer I (0.1 M NaCl + 0.05 M Na2HPO4 + 0.05 M
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NaH2PO4). First hybridization step followed. 10 µL of adenines containing oligonucleotide
and 10 µL of hybridization buffer (0.1 M phosphate buffer, 0.6 M guanidine thiocyanate,
0.15M Tris) were added in each well and then the plate was incubated for 30 minutes, at 37
°C, and 450 rpm). This procedure was followed by three times repeated washing with 100 µL
of phosphate buffer I. The second hybridization step included addition of 10 µL of
hybridization buffer and 10 µL of thiol modified DNA fragment to each well. The incubation
followed for 30 minutes, at 37 °C, and 450 rpm. Another washing with 100 µL of phosphate
buffer I followed and subsequently 30 µL of elution solution composed of phosphate buffer II
(0.2 M NaCl + 0.1 M Na2HPO4 + 0.1 M NaH2PO4) was added into each well and plate was
incubated for 5 min, at 85 °C and 450 rpm). Finally, the force of external magnetic field was
used to remove undesired impurities and resulting magnetic product was prepared for
analyses.
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