lipman03s - Supporting Online Material for Single Molecule...

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Unformatted text preview: Supporting Online Material for Single Molecule Measurement of Protein Folding Kinetics Everett A. Lipman, 1 , 2 * Benjamin Schuler, 1 , 3 * Olgica Bakajin 4 , William A. Eaton 1 1 NIDDK Laboratory of Chemical Physics, NIH Building 5 Room 116, Bethesda, MD 20892-0520, USA 2 Present address: Department of Physics, University of California, Santa Barbara, CA 93106-9530. 3 Physikalische Biochemie, Universitat Potsdam, Germany 4 Biosecurity Nanosciences Laboratory, CMS, Lawrence Livermore National Laboratory * These authors contributed equally to this work. To whom correspondence should be addressed; E-mail: eaton@helix.nih.gov. Materials and Methods Protein Figure S1 illustrates the dye-labeled cold shock protein (Csp) from Thermotoga maritima . Microfluidic Mixer A pattern of channels (Fig. 1) was cut into the surface of a silicon wafer using reactive ion etching. The channel depths varied from 8 m to 50 m, and the widths were between 5 m and 50 m. The observation channel, at one end of which the solutions were mixed, was 1 8 m deep, 50 m wide, and 10 mm long. Hash marks were etched adjacent to the observation channel at 50 m intervals so that the distance from the mixing region could be accurately determined. The concentration profiles shown in Figure 1 were modeled using pressure-determined steady-state laminar flow and sample and denaturant diffusion constants of 10- 6 cm 2 /s (= 0.1 m 2 /ms) and 10- 5 cm 2 /s (= 1 m 2 /ms), respectively. Computations were done using CFD-ACE+ software (CFDRC, Huntsville, AL). Fluid entry to and exit from the mixer took place through four pyramidal holes, etched (using KOH) through the silicon wafer at the ends of the three inlet channels and the outlet channel....
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This note was uploaded on 10/29/2009 for the course PHYS 0000 taught by Professor Many during the Spring '09 term at TN State.

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lipman03s - Supporting Online Material for Single Molecule...

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