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Biochemistry JoVE (Open Access) Citable · DOI

Fine-tuning the Size and Minimizing the Noise of Solid-state Nanopores

DOI: 10.3791/51081-v
What you'll learn
  • ✓ Control solid-state nanopore diameter with subnanometer precision using high electric field pulses
  • ✓ Characterize nanopore electrical noise and optimize signal quality in situ
  • ✓ Prepare nanopores for biomolecular translocation experiments using standard laboratory equipment
Protocol

Biopharma Insights A methodology for preparing solid-state nanopores in solution for biomolecular translocation experiments is presented. By applying short pulses of high electric fields, the nanopore diameter can be controllably enlarged with subnanometer precision and its electrical noise characteristics significantly improved. This procedure is performed in situ using standard laboratory equipment under experimental conditions.

Difficulty
advanced
Total time
~2–4 hours per nanopore (conditioning, enlargement, and characterization)

Steps

1
Characterize solid-state nanopore electrical properties

Measure baseline nanopore diameter and electrical noise characteristics using standard electrolytic measurements before modification.

▶ 01:36
2
Condition nanopores with high electric field pulses

Apply short pulses of high electric fields to stabilize nanopore structure and reduce electrical noise.

▶ 03:13
3
Enlarge nanopores using controlled electric field pulses

Incrementally increase nanopore diameter with subnanometer precision by applying calibrated high electric field pulses in situ.

▶ 04:25
4
Perform DNA translocation validation experiment

Test prepared nanopore functionality by conducting DNA translocation experiments to confirm usability for biomolecular analysis.

▶ 06:41
5
Evaluate nanopore properties and performance results

Analyze effects of high electric field conditioning and enlargement on nanopore diameter, noise characteristics, and translocation signal quality.

▶ 08:02
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