Label-Free Optical Detection of DNA Polymerase Docking on Solid-State Nanopore Arrays

• Integration of motor enzymes with biological nanopores has enabled commercial DNA sequencing, yet studies of the similar principle applying to solid-state nanopores are limited. Here, we demonstrate the label-free optical detection of phi29 DNA polymerase (DNAP) docking onto truncated-pyramidal nanopores (TPP). Atomic layer deposition of hafnium oxide is employed to shrink the pore opening size of original silicon (Si) TPP to sub-10 nm. Ionic current measurements of single TPP of an opening size comparable to DNAP show that polymerase-DNA complexes can temporally dock onto the TPP with a certain docking orientation, while the majority are translocation events. A label-free optical detection method using Ca2+ sensitive dye is employed to detect the docking of DNAP on 5-by-5 nanopore arrays. The results of the proof-of-concept experiment show that this label-free detection strategy is capable of accessing the docking events of DNAP on solid-state nanopore arrays.

Subject headings

TEKNIK OCH TEKNOLOGIER  -- Elektroteknik och elektronik -- Annan elektroteknik och elektronik (hsv//swe)

ENGINEERING AND TECHNOLOGY  -- Electrical Engineering, Electronic Engineering, Information Engineering -- Other Electrical Engineering, Electronic Engineering, Information Engineering (hsv//eng)

Publication and Content Type

vet (subject category)

ovr (subject category)