Method and materials for isolation of nucleic acid materials

Abstract

A method for nucleic acid isolation comprising: receiving a binding moiety solution within a process chamber; mixing the binding moiety solution with a biological sample, within the process chamber, in order to produce a moiety-sample mixture; incubating the moiety-sample mixture during a time window, thereby producing a solution comprising a set of moiety-bound nucleic acid particles and a waste volume; separating the set of moiety-bound nucleic acid particles from the waste volume; washing the set of moiety-bound nucleic acid particles; and releasing a nucleic acid sample from the set of moiety-bound nucleic acid particles. The method preferably utilizes a binding moiety comprising at least one of poly(allylamine) and polypropylenimine tetramine dendrimer, both of which reversibly bind and unbind to nucleic acids based upon environmental pH.

What is claimed is: 
     
         1 . A method for nucleic acid isolation from a biological sample, comprising:
 incubating affinity particles with the biological sample to reversibly bind nucleic acid material of the biological sample to the affinity particles, thereby producing affinity-bound nucleic acid particles in a sample mixture;   occluding a fluidic pathway at a first subset of a set of occlusion positions; and   separating the affinity-bound nucleic acid particles from the sample mixture, within the fluidic pathway.   
     
     
         2 . The method of  claim 1 , wherein occluding the fluidic pathway comprises displacing pins at a cartridge comprising the fluidic pathway. 
     
     
         3 . The method of  claim 2 , wherein displacing the pins comprises linearly displacing a cam card comprising a set of hills and valleys, relative to the set of pins, to a position where at least one pin of the pins transitions between a first position and a second position in a manner that occludes an occlusion position of the first subset of occlusion positions. 
     
     
         4 . The method of  claim 1 , further comprising:
 washing the affinity-bound nucleic acid particles, within the fluidic pathway; and   releasing a nucleic acid sample from the affinity-bound nucleic acid particles with an elution solution, within the fluidic pathway, wherein the nucleic acid sample comprises the nucleic acid material.   
     
     
         5 . The method of  claim 4 , wherein washing the affinity-bound nucleic acid particles is in coordination with occluding the fluidic pathway at a second subset of the set of occlusion positions. 
     
     
         6 . The method of  claim 5 , wherein releasing the nucleic acid sample comprises releasing the nucleic acid sample with the elution solution in coordination with occluding the fluidic pathway at a third subset of the set of occlusion positions, wherein the elution solution is characterized by a pH that is greater than pH 10, thereby inducing a pH shift for releasing the nucleic acid sample. 
     
     
         7 . The method of  claim 6 , further comprising spiking the nucleic acid sample with a process control comprising a set of primers and probes selected based on the nucleic acid material, within the fluidic pathway. 
     
     
         8 . The method of  claim 4 , further comprising:
 defining a fluidic path to a diagnostic chamber upon occluding the fluidic pathway at a second subset of the set of occlusion positions; and   delivering the nucleic acid sample to the diagnostic chamber through the fluidic path.   
     
     
         9 . The method of  claim 4 , wherein the affinity particles comprise microparticles amide-bonded to at least one of Poly(allylamine) (PAA) of molecular weight (MW)<40,000 Da and Polypropylenimine tetramine dendrimer (DABAM) Generation 1, and wherein releasing the nucleic acid sample comprises releasing the nucleic acid material from the at least one of the PAA of MW<40,000 Da and the DABAM Generation 1 of the affinity-bound nucleic acid particles. 
     
     
         10 . The method of  claim 1 , wherein occluding the fluidic pathway at the first subset of occlusion positions defines a fluidic path through a magnetic field, and wherein separating the affinity-bound nucleic acid particles comprises capturing the affinity-bound nucleic acid particles within the magnetic field, and delivering a waste volume of the sample mixture to a waste chamber coupled to the fluidic pathway. 
     
     
         11 . The method of  claim 10 , further comprising aspirating the nucleic acid sample within the magnetic field from the fluidic pathway; combining the nucleic acid sample with process reagents to produce a processed nucleic acid sample; and delivering the processed nucleic acid sample to a diagnostic chamber coupled to the fluidic pathway.