Low-Volume Sequencing System and Method of Use

Abstract

Various embodiments of a low-volume sequencing system are provided herein. The system can include a low-volume flowcell having at least one reaction chamber of a defined volume (e.g., less than about 100 μl). The system can also include an automated reagent delivery mechanism configured to reversibly couple with the inlet port corresponding to a target reaction chamber thereby placing allowing for reagent to be accurately moved from a storage container to the reaction chamber with minimal reagent waste. The flowcells can include a plurality of reaction chambers (e.g., 6) thereby allowing for parallel analysis of multiple samples. Various methods of analyzing a biomolecule are also provided herein.

What is claimed is: 
     
         1 . A low-volume sequencing system, comprising:
 a low-volume flowcell having at least one reaction chamber of a defined volume and at least one fluid transfer port formed therein; and   an automated reagent delivery mechanism having a fluid dispenser with an internal compartment configured to retain a selected amount of reagent, the fluid dispenser having a distal portion configured to reversibly couple with the inlet port thereby placing the internal compartment into fluid communication with the at least one reaction chamber, and further configured to dispense to the reaction chamber a volume substantially equal to the defined volume of the reaction chamber.   
     
     
         2 . The system of  claim 1 , wherein the delivery mechanism is configured to retain a volume of reagent substantially equal to the volume of the reaction chamber. 
     
     
         3 . The system of  claim 1 , further comprising a controller in communication with the delivery mechanism, and configured to couple or decouple the delivery mechanism with the fluid transfer port, and further configured to dispense reagent to the reaction chamber. 
     
     
         4 . The system of  claim 1 , wherein the defined volume of the reaction chamber is less than about 100 μl. 
     
     
         5 . The system of  claim 1 , wherein the flowcell includes a plurality of reaction chambers, each reaction chamber having a discrete volume. 
     
     
         6 . The system of  claim 5 , wherein each reaction chamber extends between at least two discrete fluid transfer ports. 
     
     
         7 . The system of  claim 5 , wherein the internal compartment of delivery mechanism is configured to retain a volume of a reagent substantially equal to a total volume of the plurality of reaction chambers. 
     
     
         8 . The system of  claim 1 , wherein at least one fluid transfer port is incorporated into a top portion of the low-volume flowcell. 
     
     
         9 . The system of  claim 1 , wherein at least one fluid transfer port is incorporated into a bottom portion of the low-volume flowcell. 
     
     
         10 . The system of  claim 1 , wherein at least a portion of the flowcell surface is configured to bind a sample. 
     
     
         11 . The system of  claim 10 , wherein the sample is a polynucleotide or the sample is a solid support configured to bind a polynucleotide or a polynucleotide. 
     
     
         12 . The system of  claim 1 , wherein an interior surface of the flowcell is configured to bind sample. 
     
     
         13 . The system of  claim 1 , wherein the reagent delivery mechanism includes a robotic assembly having x-, y-, and z-functionality. 
     
     
         14 . The system of  claim 1 , wherein the low-volume flowcell includes a plurality of discrete reaction chambers. 
     
     
         15 . The system of  claim 1 , further comprising a reagent storage container housing a plurality of reagents. 
     
     
         16 . A low-volume flowcell, comprising:
 a substrate having at least one reaction chamber of a defined volume extending between an inlet port and an outlet port, the inlet port being configured to reversibly couple with an automated reagent delivery mechanism which is configured to deliver a pre-determined amount of reagent to the reaction chamber, the reaction chamber being sized and configured to minimize the pre-determined amount of reagent required to effect a desired result.   
     
     
         17 . The low-volume flowcell of  claim 16 , wherein the defined volume is between about 15 μl and about 35 μl. 
     
     
         18 . The low-volume flowcell of  claim 16 , wherein the defined volume is between about 10 μl and about 40 μl. 
     
     
         19 . The low-volume flowcell of  claim 16 , wherein the defined volume is about 25 μl. 
     
     
         20 . A method of analyzing a biomolecule, comprising:
 providing an automated reagent delivery mechanism configured to withdraw a pre-determined volume of a reagent from a reagent storage container;   withdrawing a pre-determined amount of the reagent from the storage container by the automated reagent delivery mechanism;   coupling a portion of the automated reagent delivery mechanism to an inlet port of a flowcell, the inlet port being in fluid communication with a reaction chamber having a defined volume;   dispensing into the reagent chamber a reagent volume substantially equal to the defined volume of the reagent chamber; and   decoupling the automated reagent delivery mechanism from the inlet port.