For details on the nanodisk structure see Supplementary Table 2 . Folding is carried out over three days (for folding program see Supplementary Table 3 ) in 1xTAE buffer containing 12.5 mM MgCl2. The DNA origamis are filtered three times at 15,000 rcf for 5 minutes (Amicon Ultra-0.5 ml Centrifugal Filters, 100,000 MWCO, Millipore) with pH 7.4 MOPS buffer containing 50 mM MOPS, 75 mM potassium acetate and 12.5 mM magnesium acetate. The sample is recovered for 3 minutes at 3,000 rcf. We purchase ZMW samples from Pacific Biosciences, where the ZMW chip is mounted on a conventional microscope slide. Each chip provides 6 different ZMW diameters ranging from 85-200 nm. The glass bottom of the ZMW chip is already functionalized with PEG-biotin to allow immobilization of neutravidin labelled molecules 46 (link). The chip is washed twice with MOPS buffer before incubation with 1 mg/ml neutravidin in MOPS buffer. After five more washing steps, the DNA origami sample is added and the binding process is controlled by confocal imaging. Once a reasonable occupancy is reached, the sample is thoroughly washed to remove unbound DNA structures. Measurements are carried out in MOPS buffer with 1% (w/v) glucose and 10% (v/v) of the oxygen scavenging system.
Amicon ultra 0.5 ml centrifugal filters 100 000 mwco
Manufactured by Merck Group
The Amicon Ultra-0.5 ml Centrifugal Filters, 100,000 MWCO are laboratory filtration devices used for the concentration and purification of macromolecules such as proteins, peptides, and nucleic acids. These filters have a molecular weight cutoff of 100,000 Daltons, allowing the effective separation of the target molecules from smaller contaminants or buffer components.
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2 protocols using amicon ultra 0.5 ml centrifugal filters 100 000 mwco
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Nanodisk DNA Origami Immobilization
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Nanodisk DNA Origami Immobilization
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For details on the nanodisk structure see Supplementary Table 2 . Folding is carried out over three days (for folding program see Supplementary Table 3 ) in 1xTAE buffer containing 12.5 mM MgCl2. The DNA origamis are filtered three times at 15,000 rcf for 5 minutes (Amicon Ultra-0.5 ml Centrifugal Filters, 100,000 MWCO, Millipore) with pH 7.4 MOPS buffer containing 50 mM MOPS, 75 mM potassium acetate and 12.5 mM magnesium acetate. The sample is recovered for 3 minutes at 3,000 rcf. We purchase ZMW samples from Pacific Biosciences, where the ZMW chip is mounted on a conventional microscope slide. Each chip provides 6 different ZMW diameters ranging from 85-200 nm. The glass bottom of the ZMW chip is already functionalized with PEG-biotin to allow immobilization of neutravidin labelled molecules 46 (link). The chip is washed twice with MOPS buffer before incubation with 1 mg/ml neutravidin in MOPS buffer. After five more washing steps, the DNA origami sample is added and the binding process is controlled by confocal imaging. Once a reasonable occupancy is reached, the sample is thoroughly washed to remove unbound DNA structures. Measurements are carried out in MOPS buffer with 1% (w/v) glucose and 10% (v/v) of the oxygen scavenging system.
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