Circular supercoiled pUC18 plasmid was purified using a PureYield™ Plasmid Miniprep kit (Promega, Madison, WI, USA) following the provided protocol with two additional wash steps. Protein–DNA complexes were obtained by adding 4.5 ng purified pUC18 plasmid in Lrp binding buffer to varying concentrations of protein in a total volume of 15 μL, followed by a 20 min incubation at 37 °C. After incubation, the prepared samples were mixed with an equal volume of nickel absorption buffer (40 mM HEPES, 10 mM NiCl2 (pH 6.74)). For each binding reaction, 10 μL was disposed onto a freshly cleaved mica disc and incubated for another 10 min to allow adsorption of the DNA on the mica. The mica surface was rinsed 5 times with water and gently dried with a stream of air. Atomic force microscopy (AFM) images were obtained using a Multimode Nanoscope IIIa AFM (Bruker, Billerica, MA, USA) operated in tapping mode in air. RTESP AFM probes (Bruker, Billerica, MA, USA) were ozone-cleaned just before use. Images were recorded at a scan rate of 1.5 Hz. Flattening of the images was performed by using NanoScope Analysis v1.5 software (Bruker, Billerica, MA, USA). 3-D images were shown with a pitch of 3°.
Rtesp afm probes
Manufactured by Bruker
Sourced in United States
The RTESP AFM probes are designed for tapping mode atomic force microscopy (AFM) measurements. They feature a rectangular cantilever and a silicon tip, providing high-resolution imaging capabilities. The probes are suitable for a variety of applications that require tapping mode AFM functionality.
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Lab products found in correlation
Nanoscope iiia multimode afm, by Bruker (1 mentions)
Nanoscope analysis v1, by Bruker (1 mentions)
Pureyield plasmid miniprep kit, by Promega (1 mentions)
D8 discover x ray diffraction, by Bruker (1 mentions)
Atomic force microscope, by Bruker (1 mentions)
S 4800 scanning electron microscope, by Hitachi (1 mentions)
2 protocols using rtesp afm probes
1
AFM Visualization of Protein-DNA Complexes
2
Characterization of ZnO Nanowires
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The quality of the resultant ZnO nanowire growth was studied with a Hitachi S-4800 scanning electron microscope (SEM) at 10 kV to obtain topographical information such as diameter, shape and density of growth. The SEM was equipped with an Oxford Instruments energy-dispersive x-ray detector (EDX) employed to determine the elemental composition of the grown samples. The EDX was also used to assess changes in the elemental composition of the seed layers with respect to UV exposure time. A Bruker D8 Discover x-ray diffraction (XRD) system, equipped with a Cu_Kα x-ray source and a Lynxeye detector, was employed to evaluate the crystallinity of the as grown ZnO nanowires. Furthermore x-ray photoelectron spectroscopy (XPS) was used to evaluate the structure and composition of the seed layers with respect to UV exposure. The XPS experiments were carried out at room temperature using the Kratos Axis Supra system equipped with a monochromatic Al Kα source having an energy of 1486.6 eV. To investigate surface morphology of the seed layers a JPK Instruments atomic force microscope (AFM) was used in AC mode using tapping mode RTESP AFM probes (Bruker, USA). Images were subsequently analysed using scanning probe image processor (SPIP) image analysis software.
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