System and methods for stretching polynucleotides
AssigneeMassachusetts Institute of Technology
InventorsGreg Randall, Patrick Doyle
Life of this patent
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Abstract
Apparatus and methods are described for achieving uniform stretching of polynucleotides in hybrid electrophoretic-gel, micro-constricting microfluidic channels. Polynucleotides in normally relaxed configurations are driven by an electric field along a microfluidic channel. The polynucleotides thread through a porous gel barrier formed in the channel and extend into a constriction where an electric field gradient exists. The combined action of the gel and field gradient acts to extend the polynucleotide configuration fully for direct linear analysis of the molecule.
Claims22
An apparatus for extending long molecules from a coiled configuration to an extended configuration comprising:
- a substrate;
- a fluidic channel defined on the substrate;
- a cover disposed on the substrate, covering at least a portion of the fluidic channel and forming a lumen;
- a sample entry portion;
- a sample exit portion;
- a constricting section defined within at least a portion of the fluidic channel;
- a porous barrier disposed proximally and upstream of the constricting section of the fluidic channel wherein pore sizes at an upstream side of the porous barrier are about equal to one persistence length of the molecules passing through the barrier, and wherein pore sizes increase to multiple persistence lengths at a downstream side of the porous barrier;
- and
- electrodes adapted to apply an electric field in a direction substantially along the length of the fluidic channel such that an electric field gradient is established within the constricting section,
- wherein the fluidic channel connects the sample entry portion to the sample exit portion.
An apparatus for extending long molecules from a coiled configuration to an extended configuration comprising:
- a constricting section defined within at least a portion of a microfluidic channel;
- a porous barrier disposed proximally and upstream of the constricting section wherein pore sizes at an upstream side of the porous barrier are about equal to one persistence length of the molecules passing through the barrier, and wherein pore sizes increase to multiple persistence lengths at a downstream side of the porous barrier;
- and
- an electric field gradient within the constricting section of the microfluidic channel, wherein the porous harrier and electric field gradient cooperate in extending the long molecules.
Dependencies are inferred from the claim text.