Nucleic acid purification

Abstract

A self-contained apparatus for isolating nucleic acid, cell lysates and cell suspensions from unprocessed samples apparatus, to be used with an instrument, includes at least one input, and: (i) a macrofluidic component, including a chamber for receiving an unprocessed sample from a collection device and at least one filled liquid purification reagent storage reservoir; and (ii) a microfluidic component in communication with the macrofluidic component through at least one microfluidic element, the microfluidic component further comprising at least one nucleic acid purification matrix; and (iii) at least one interface port to a drive mechanism on the instrument for driving said liquid purification reagent, through the microfluidic element and the nucleic acid purification matrix, wherein the only inputs to the apparatus are through the chamber and the interface port to the drive mechanism.

1 - 9 . (canceled) 
     
     
         10 . A self-contained apparatus for isolating nucleic acid from at least one unprocessed sample, said apparatus to be used with an instrument,
 said apparatus consisting of a macrofluidic component, a microfluidic component, at least one drive line, and two inputs, a first input for receiving the at least one unprocessed sample from a collection device, and a second input for interfacing at least one pneumatic drive mechanism on said instrument at, at least one port;
 (a) said macrofluidic component comprising: (i) at least one sample chamber, each of said at least one sample chamber composed of said first input for receiving said at least one unprocessed sample from the collection device; (ii) at least one, but no less than all reagent storage chambers necessary for isolating said nucleic acid from said at least one unprocessed sample, each of said reagent storage chambers pre-filled with at least one necessary reagent, including, at least one lysis reagent storage chamber pre-filled with lysis reagent; 
 (b) said microfluidic component in fluid communication with said microfluidic component; 
 (c) said second input composed of at least one pneumatic drive mechanism interface port for connection to said at least one drive mechanism on said instrument configured to chaotically bubble said at least one lysis reagent in the at least one lysis reagent storage chamber, resulting in turbulent flow that releases nucleic acid from each of said at least one unprocessed sample. 
   
     
     
         11 . The apparatus of  claim 10  wherein the macrofluidic component further comprises: an elution reagent storage chamber pre-filled with elution reagent, and wherein said microfluidic component further comprises at least one nucleic acid purification matrix, and wherein said pneumatic drive mechanism on said instrument is configured to bubble said elution reagent and to drive said bubbled elution reagent through said nucleic acid purification matrix. 
     
     
         12 . The apparatus of  claim 10  wherein the nucleic acid purification matrix is selected from the group consisting of silica membranes, silica beads, silica magnetic beads, ion exchange resins, and ion exchange beads. 
     
     
         13 . The apparatus of  claim 10  wherein the unprocessed sample is received from a collection device having a swab head which is subjected to chaotic bubbling to mediate cell lysis from the swab head prior to the addition of lysis reagent. 
     
     
         14 . The apparatus of  claim 11  wherein the swab head is substantially cotton. 
     
     
         15 . The apparatus of  claim 10  wherein said microfluidic component further comprises at least one particulate filter. 
     
     
         16 . The apparatus of  claim 10  wherein said microfluidic component further comprises at least one microfluidic element selected from the group consisting of: channels, reservoirs, active valves, passive valves, pneumatically actuated valves, reaction chambers, mixing chambers, venting elements, access holes, pumps, metering elements, mixing elements, heating elements, magnetic elements, reaction chambers, filtration elements, purification elements, drive lines and actuation lines. 
     
     
         17 . A self-contained apparatus for isolating nucleic acid from at least one unprocessed sample, said apparatus to be used with an instrument, said apparatus comprising at least one input, and
 (a) a macrofluidic component comprising:   at least one chamber for receiving said at least one unprocessed sample; and   at least one filled reagent storage reservoir containing a substantially isotonic reagent;   (b) a microfluidic component in communication with said macrofluidic component via at least one microfluidic element, said microfluidic component further comprising at least one nucleic acid purification matrix,   (c) a pneumatic drive mechanism interface port for interfacing said instrument;   wherein the only inputs to said apparatus are via said at least one chamber and said drive mechanism interface port, and wherein upon instruction from said instrument, air is driven into said at least one filled reagent storage reservoir to chaotically bubble said substantially isotonic reagent with said at least one unprocessed sample, resulting in turbulent flow that releases nucleic acid from said sample, and wherein released nucleic acid is thereafter bubbled through said nucleic acid purification matrix, resulting in isolated nucleic acid in at least one microfluidic element.   
     
     
         18 . The apparatus of  claim 17 , wherein said at least one microfluidic element is selected from a group consisting of channels, reservoirs, active valves, passive valves, pneumatically actuated valves, reaction chambers, mixing chambers, venting elements, access holes, pumps, metering elements, mixing elements, heating elements, magnetic elements, reaction chambers, filtration elements, purification elements, drive lines and actuation lines. 
     
     
         19 . The apparatus of  claim 17  wherein the nucleic acid purification matrix is selected from the group consisting of silica membranes, silica beads, silica magnetic beads, ion exchange resins, and ion exchange beads. 
     
     
         20 . The apparatus of  claim 17  wherein the unprocessed sample is received from a collection device having a swab head which is subjected to chaotic bubbling to mediate cell lysis from the swab head prior to the addition of substantially isotonic reagent. 
     
     
         21 . The apparatus of  claim 17  wherein the swab head is substantially cotton. 
     
     
         22 . A self-contained apparatus for isolating nucleic acid from at least one unprocessed sample, said apparatus to be used with an instrument, said apparatus consisting of a macrofluidic component, a microfluidic component, at least one drive line, and two inputs, a first input for receiving the at least one unprocessed sample from a collection device, and a second input for interfacing at least one pneumatic drive mechanism on said instrument at, at least one pneumatic drive mechanism interface port:
 (a) said macrofluidic component comprising: (i) at least one sample chamber, each of said at least one sample chamber composed of said first input for receiving said at least one unprocessed sample from the collection device; (ii) at least one, but no less than all reagent storage chambers necessary for isolating said nucleic acid from said at least one unprocessed sample, each of said reagent storage chambers pre-filled with at least one necessary reagent, including, at least one lysis reagent storage chambers pre-filled with lysis reagent; a wash reagent storage chamber pre-filled with wash reagent; and an elution reagent storage chamber pre-filled with elution reagent;   (b) said microfluidic component comprising at least one nucleic acid purification matrix, said microfluidic component in communication with said macrofluidic component;   (c) said second input composed of at least one pneumatic drive mechanism interface port for connection to said at least one pneumatic drive mechanism on said instrument; and configured to chaotically bubble said at least one lysis reagent pre-filled in the at least one lysis reagent storage chamber, and subsequentially to chaotically bubble said elution reagent; and to drive said bubbled elution reagent through said at least one nucleic acid purification matrix.   
     
     
         23 . The apparatus of  claim 22 , wherein said at least one microfluidic element is selected from a group consisting of channels, reservoirs, active valves, passive valves, pneumatically actuated valves, reaction chambers, mixing chambers, venting elements, access holes, pumps, metering elements, mixing elements, heating elements, magnetic elements, reaction chambers, filtration elements, purification elements, drive lines and actuation lines. 
     
     
         24 . The apparatus of  claim 22 , wherein a first lysis reagent storage chamber of said at least two lysis reagent storage chambers is pre-filled with guanidine and a second lysis reagent storage chamber of said at least two lysis reagent storage chambers is pre-filled with ethyl alcohol and said wash reagent storage chamber is pre-filled with an ethanol based wash reagent. 
     
     
         25 . The apparatus of  claim 22 , wherein an aggregate fluid volume of the at least one sample chamber, the at least two lysis reagent storage chambers, the wash reagent storage chamber, and the elution reagent storage chamber is between about 1 and 1000 mL. 
     
     
         26 . A self-contained apparatus for isolating nucleic acid from at least one unprocessed sample, said apparatus to be used with an instrument,
 said apparatus consisting of a macrofluidic component, a microfluidic component, at least one drive line, and two inputs, a first input for receiving the at least one unprocessed sample from a collection device, and a second input for interfacing at least one drive mechanism on said instrument at, at least one port:   said macrofluidic component comprising: (i) at least one sample chamber, each of said at least one sample chamber composed of said first input for receiving said at least one unprocessed sample from the collection device; (ii) at least one, but no less than all reagent storage chambers necessary for isolating said nucleic acid from said at least one unprocessed sample, each of said reagent storage chambers pre-filled with at least one necessary reagent, including, at least one lysis reagent storage chamber pre-filled with lysis reagent, which, upon release of lysis reagent is capable of functioning as a lysis/waste chamber; a wash reagent storage chamber pre-filled with wash reagent; and an elution reagent storage chamber pre-filled with elution reagent and an elution reagent homogenization chamber in fluid communication with said microfluidic component;   said microfluidic component comprising at least one nucleic acid purification matrix, said microfluidic component in communication with said macrofluidic component;   said second input composed of at least one drive mechanism interface port for connection to said at least one drive mechanism on said instrument;   whereupon in use, unprocessed or partially processed sample is pneumatically driven into the lysis chamber prefilled with lysis reagent; air is pneumatically driven into the lysis chamber to effect chaotic bubbling of the lysate within the lysis chamber; ethanol is driven into the lysate chamber by continued chaotic bubbling, whereupon all the lysate and ethanol mixture is pneumatically driven into the lysis chamber; and thereafter through the purification matrix and back into the lysis chamber which now functions as a lysis/waste chamber; and thereupon wash solution is driven from the wash reservoir through the purification membrane and into the lysis/waste chamber; and elution reagent is bubbled from the elution reagent reservoir through the purification membrane to the elution reagent homogenization chamber, resulting in homogenized purified DNA solution in said eluate homogenization chamber.   
     
     
         27 . The apparatus of  claim 26  wherein the unprocessed sample is received from a collection device having a swab head which is subjected to chaotic bubbling to mediate cell lysis from the swab head prior to the addition of lysis reagent. 
     
     
         28 . The apparatus of  claim 26  wherein the swab head is substantially cotton.