Hybrid pore formation by directed insertion of α-haemolysin into solid-state nanopores

Most experiments on nanopores have concentrated on the pore-forming protein alpha-haemolysin (alpha HL)(1) and on artificial pores in solid-state membranes(2). While biological pores offer an atomically precise structure(3) and the potential for genetic engineering(4), solid-state nanopores offer durability, size and shape control(5), and are also better suited for integration into wafer-scale devices. However, each system has significant limitations: alpha HL is difficult to integrate because it relies on delicate lipid bilayers for mechanical support, and the fabrication of solid-state nanopores with precise dimensions remains challenging. Here we show that these limitations may be overcome by inserting a single alpha HL pore into a solid-state nanopore. A double-stranded DNA attached to the protein pore is threaded into a solid-state nanopore by electrophoretic translocation. Protein insertion is observed in 30-40% of our attempts, and translocation of single-stranded DNA demonstrates that the hybrid nanopore remains functional. The hybrid structure offers a platform to create wafer-scale device arrays for genomic analysis, including sequencing(6).