Variable Valve Apparatus and Methods

Abstract

Sample processing devices with variable valve structures and methods of using the same are disclosed. The valve structures allow for removal of selected portions of the sample material located within the process chamber. Removal of the selected portions is achieved by forming an opening in a valve septum at a desired location. The valve septums may be large enough to allow for adjustment of the location of the opening based on the characteristics of the sample material in the process chamber. If the sample processing device is rotated after the opening is formed, the selected portion of the material located closer to the axis of rotation exits the process chamber through the opening. The remainder of the sample material cannot exit through the opening because it is located farther from the axis of rotation than the opening.

1 . A method of amplifying nucleic acid, the method comprising: 
 providing a device comprising a process array that comprises process chambers defining volumes for containing sample material and a conduit connecting the process chambers;    delivering sample material comprising nucleic acid-containing material to a process chamber;    contacting the nucleic acid-containing material with a lysing reagent; and    transferring the sample material and the lysing reagent to a different process chamber through the conduit by rotating the device, wherein the sample material contacts amplification reagents located in the different process chamber; and    amplifying nucleic acids in the sample material in the different process chamber.    
     
     
         2 . The method of  claim 1  further comprising heating the sample material to a temperature at which the lysing reagent is inactivated prior to transferring the sample material.  
     
     
         3 . The method of  claim 1  wherein the nucleic acid-containing material comprises cells.  
     
     
         4 . The method of  claim 3  wherein the cells are eukaryotic.  
     
     
         5 . The method of  claim 4  wherein the eukaryotic cells comprise tissue cells.  
     
     
         6 . The method of  claim 5  wherein the lysing reagent comprises a proteinase.  
     
     
         7 . The method of  claim 6  wherein the proteinase is proteinase K.  
     
     
         8 . The method of  claim 3  wherein the cells are prokaryotic.  
     
     
         9 . The method of  claim 8  wherein the prokaryotic cells comprise bacteria.  
     
     
         10 . The method of  claim 9  wherein the lysing reagent comprises an enzyme or a base.  
     
     
         11 . The method of  claim 3  wherein the nucleic acid-containing material comprises bacteria.  
     
     
         12 . The method of  claim 11  wherein the bacteria are gram positive bacteria.  
     
     
         13 . The method of  claim 12  wherein the lysing reagent comprises a lysozyme.  
     
     
         14 . The method of  claim 1  wherein contacting the nucleic acid-containing material with a lysing reagent comprises using at least two lysing reagents.  
     
     
         15 . The method of  claim 1  wherein the nucleic acid-containing material comprises a virus.  
     
     
         16 . The method of  claim 1  wherein contacting the nucleic acid-containing material with the lysing reagent occurs before the sample material is delivered to the valved process chamber.  
     
     
         17 . A method of amplifying nucleic acid, the method comprising: 
 providing sample material comprising nucleic acid-containing material;    providing a device comprising a process array that comprises process chambers defining volumes for containing sample material and a conduit connecting the process chambers;    concentrating the nucleic acid-containing material;    contacting the concentrated nucleic acid-containing material with a lysing reagent in a process chamber; and    transferring the sample material and the lysing reagent to a different process chamber through the conduit by rotating the device, wherein the sample material contacts amplification reagents located in the different process chamber; and    amplifying nucleic acids in the sample material in the different process chamber.    
     
     
         18 . The method of  claim 17  wherein concentrating the nucleic acid-containing material comprises capturing the nucleic acid-containing material with functionalized beads.  
     
     
         19 . The method of  claim 18  further comprising heating the sample material to a temperature at which the lysing reagent is inactivated prior to transferring the sample material.  
     
     
         20 . The method of  claim 17  wherein the nucleic acid-containing material comprises cells.  
     
     
         21 . The method of  claim 20  wherein the cells are eukaryotic.  
     
     
         22 . The method of  claim 21  wherein the eukaryotic cells comprise tissue cells.  
     
     
         23 . The method of  claim 20  wherein the cells are prokaryotic.  
     
     
         24 . The method of  claim 23  wherein the prokaryotic cells comprise bacteria.  
     
     
         25 . The method of  claim 17  wherein the lysing reagent comprises a proteinase.  
     
     
         26 . The method of  claim 25  wherein the proteinase comprises proteinase K.  
     
     
         27 . The method of  claim 17  wherein the lysing reagent comprises an enzyme or a base.  
     
     
         28 . The method of  claim 17  wherein the nucleic acid-containing material comprises bacteria.  
     
     
         29 . The method of  claim 28  wherein the bacteria comprise gram positive bacteria.  
     
     
         30 . The method of  claim 17  wherein the nucleic acid-containing material comprises a virus.  
     
     
         31 . A method of amplifying nucleic acid, the method comprising: 
 providing sample material comprising nucleic acid-containing material;    providing a device comprising a process array that comprises process chambers defining volumes for containing sample material and a conduit connecting the process chambers;    contacting the nucleic acid-containing material with a lysing reagent in a process chamber;    capturing the nucleic acid with functionalized beads;    eluting the nucleic acid; and    transferring the nucleic acid to a different process chamber through a conduit by rotating the device, wherein the nucleic acid contacts amplification reagents located in the different process chamber; and    amplifying nucleic acid in the different process chamber.    
     
     
         32 . A method of amplifying nucleic acid, the method comprising: 
 providing sample material comprising nucleic acid-containing material;    providing a device comprising a process array, wherein the process array comprises process chambers defining volumes for containing sample material and a conduit connecting the process chambers;    concentrating the nucleic acid-containing material;    lysing the concentrated nucleic acid-containing material;    denaturing proteins within the sample material in a process chamber;    transferring the sample material to a different process chamber through the conduit after denaturing the proteins, wherein the transferring comprises rotating the device, and wherein the sample material contacts amplification reagents located in the different process chamber; and    amplifying nucleic acids in the sample material in the different process chamber.    
     
     
         33 . The method of  claim 32  wherein denaturing occurs after lysing the nucleic acid-containing material.  
     
     
         34 . The method of  claim 33  wherein the denaturing is performed while the sample material is in the process chamber.  
     
     
         35 . The method of  claim 32  wherein the denaturing comprises heating the sample material.  
     
     
         36 . The method of  claim 32  wherein the lysing is performed by heating the sample material.  
     
     
         37 . The method of  claim 32  wherein the lysing is performed using a lysing reagent.  
     
     
         38 . The method of  claim 17  wherein the lysing agent comprises a lysozyme.