Nanopore/nanowell electrode enabled exonuclease sequencing

A nanopore or a nanowell device comprising:

• a substrate comprising a chamber comprising a proximal opening, a distal opening, sidewalls and an interior volume;
• a first seal over the proximal opening of the chamber and a second seal over the distal opening of the chamber;
• a transmembrane channel in the first seal oriented with a channel entrance exposed to a solution outside the nanopore electrode device (cis) and a channel exit exposed to a solution inside the nanopore electrode device (trans);
• an enzyme attached to an interior surface of the chamber at a distance from the first seal greater than 100 nm to about 2000 nm;
• and
• a component configured to detect an interaction between a molecule and the transmembrane channel.

A method for determining the sequence of a polymer, comprising:

• a) electrophoretically and/or electroosmotically driving a polymer from a bulk solution through a transmembrane channel disposed in a membrane into a confined volume of a chamber;
• whereby the polymer contacts an enzyme in the confined volume of the chamber, under conditions in which the enzyme cleaves monomeric units from the polymer;
• b) electrophoretically and/or electroosmotically driving monomeric units in the order which they are sequentially cleaved from the polymer by the enzyme out of the confined volume of the chamber through the transmembrane channel;
• and
• c) determining the identity of each of the monomeric units based on its current signature, translocation time, and/or associated current noise level modulation as the monomeric unit translocates through the transmembrane channel, thereby determining the sequence of the polymer.

The method of claim 18, wherein the transmembrane channel comprises:

• (a) a mutated, engineered, chemically modified transmembrane channel,
• (b) a mutant form transmembrane channel comprising a modification of the transmembrane channel;
• or
• (c) a solid state nanopore.