ERASE: a novel surface reconditioning strategy for single-molecule experiments

• Published in: Nucleic acids research 2019-02, Vol.47 (3), p.e14-e14
• Main Authors: Broadwater, D W Bo, Altman, Roger B, Blanchard, Scott C, Kim, Harold D
• Format: Article
• Language: English
• Subjects: Carbocyanines; Fluorescent Dyes; Methods Online; Nucleic Acid Hybridization; Oligodeoxyribonucleotides - chemistry; Single Molecule Imaging
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/gky1168

Abstract While surface-based single-molecule experiments have revolutionized our understanding of biology and biomolecules, the workflow in preparing for such experiments, especially surface cleaning and functionalization, remains labor-intensive and time-consuming. Even worse, meticulously assembled flow channels can be used only once for most experiments. A reusable surface would thus dramatically increase productivity and efficiency of single-molecule experiments. In this paper, we report a novel surface reconditioning strategy termed ERASE (Epitaxial Removal Aided by Strand Exchange) that allows a single flow cell to be used for vast repetition of single-molecule experiments. In this method, biomolecules immobilized to the surface through a nucleic acid duplex are liberated when a competing DNA strand disrupts the duplex via toehold-mediated strand displacement. We demonstrate the wide-range applicability of this method with various common surface preparation techniques, fluorescent dyes, and biomolecules including the bacterial ribosome. Beyond time and cost savings, we also show ERASE can assort molecules based on a nucleic acid barcode sequence, thus allowing experiments on different molecules in parallel. Our method increases the utility of prepared surfaces and is a significant improvement to the current single-use paradigm.