Macromolecule delivery to nanowells

Abstract

Provided herein is technology relating to depositing and/or placing a macromolecule at a desired site for an assay and particularly, but not exclusively, to methods and systems for transporting a macromolecule such as a protein, a nucleic acid, or a protein:nucleic acid complex to an assay site, such as the bottom of a nanopore, a nanowell, or a zero mode waveguide.

We claim: 
     
         1 . A method for delivering a macromolecule to an assay site, wherein the method comprises:
 1) maintaining an end of a transport guide at the assay site;   2) providing a molecular motor adapted for binding the transport guide and moving along the transport guide; and   3) linking the macromolecule to the molecular motor.   
     
     
         2 . The method of  claim 1 , wherein the assay site is in a zero-mode waveguide or in a nanowell. 
     
     
         3 . The method of  claim 1 , wherein the assay site comprises an anchor to maintain the macromolecule at the assay site. 
     
     
         4 . The method of  claim 1 , wherein:
 a) the transport guide is a microtubule and the molecular motor is selected from the group consisting of a kinesin and a dynein; or   b) the transport guide is an actin filament and the molecular motor is a myosin.   
     
     
         5 . The method of  claim 1 , wherein the macromolecule comprises a DNA polymerase and/or a DNA. 
     
     
         6 . The method of  claim 1 , further comprising providing a DNA polymerase at the assay site. 
     
     
         7 . The method of  claim 1 , wherein maintaining the end of the transport guide at the assay site comprises attaching the end of the transport guide to the assay site. 
     
     
         8 . The method of  claim 1 , wherein the linking comprises providing a domain selected from the group consisting of a myosin binding domain and a microtubule associated protein binding domain. 
     
     
         9 . The method of  claim 1 , wherein the transport guide is stabilized with a composition selected from the group consisting of a phalloidin and a paclitaxel. 
     
     
         10 . The method of  claim 1 , wherein the transport guide is disassembled and/or destabilized by a cytochalasin. 
     
     
         11 . The method of  claim 1 , wherein the linking is a covalent interaction with the molecular motor. 
     
     
         12 . The method of  claim 1 , wherein the linking is a non-covalent interaction with the molecular motor. 
     
     
         13 . The method of  claim 1 , wherein the linking comprises a streptavidin and/or a biotin.