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A DNA-based molecular probe for optically reporting cellular traction forces.

A DNA-based molecular probe for optically reporting cellular traction forces.

Nature methods (2014-10-13)

Brandon L Blakely, Christoph E Dumelin, Britta Trappmann, Lynn M McGregor, Colin K Choi, Peter C Anthony, Van K Duesterberg, Brendon M Baker, Steven M Block, David R Liu, Christopher S Chen

ABSTRACT

We developed molecular tension probes (TPs) that report traction forces of adherent cells with high spatial resolution, can in principle be linked to virtually any surface, and obviate monitoring deformations of elastic substrates. TPs consist of DNA hairpins conjugated to fluorophore-quencher pairs that unfold and fluoresce when subjected to specific forces. We applied TPs to reveal that cellular traction forces are heterogeneous within focal adhesions and localized at their distal edges.

MATERIALS

Product Number

Brand

Product Description

Riboflavin, European Pharmacopoeia (EP) Reference Standard

Millipore

Urea solution, suitable for microbiology, 40% in H₂O

Sigma-Aldrich

Methanol, suitable for NMR (reference standard)

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Methanol, purification grade, 99.8%

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DL-Dithiothreitol solution, 1 M in H₂O

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Magnesium chloride solution, 0.1 M

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Acetic acid, ≥99.5%, FCC, FG

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Propylamine, ≥99%

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Acetic acid-¹²C₂, 99.9 atom % ¹²C

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Propylamine, analytical standard

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Formamide, Vetec^™, reagent grade, 98%

USP

Riboflavin, United States Pharmacopeia (USP) Reference Standard

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Formamide, ≥99.5% (GC), BioReagent, Molecular Biology

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Urea solution, 40 % (w/v) in H₂O

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Magnesium chloride solution, PCR Reagent, 25 mM MgCI₂solution for PCR

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Ethanol standards 10% (v/v), 10 % (v/v) in H₂O, analytical standard

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(−)-Riboflavin, meets USP testing specifications

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Magnesium chloride solution, Molecular Biology, 1.00 M±0.01 M

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Ethanol Fixative 80% v/v, suitable for fixing solution (blood films)

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(−)-Riboflavin, from Eremothecium ashbyii, ≥98%

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Acetic acid, suitable for luminescence, BioUltra, ≥99.5% (GC)

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Magnesium chloride solution, BioUltra, Molecular Biology, ~0.025 M in H₂O

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Magnesium chloride solution, BioUltra, Molecular Biology, ~1 M in H₂O

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Propylamine, 98%

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Acetic acid, analytical standard

Supelco

Methanol, analytical standard

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Urea solution, BioUltra, ~8 M in H₂O

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DL-Dithiothreitol solution, BioUltra, Molecular Biology, ~1 M in H₂O

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Methanol, anhydrous, 99.8%

Sigma-Aldrich

Urea-¹²C, 99.9 atom % ¹²C