Kits for Isolating Nucleic Acids Using Multifunctional Group-Coated Solid Phase Carriers

Abstract

The present invention is directed to a method of isolating a target species (e.g., target nucleic acid species) from a mixture. In the methods of the present invention, the mixture is combined with solid phase carriers having a surface comprising multiple functional groups one of which reversibly and selectively binds the target species. In a particular embodiment, the mixture is combined with solid phase carriers having a first functional group which reversibly binds nucleic acids and a second functional group which selectively and reversibly binds the target nucleic acid species, thereby producing a first combination. The first combination is maintained under conditions appropriate for binding of the nucleic acids to the first functional group and binding of the target nucleic acid species to the second functional group. The solid phase carriers are separated from the first combination, and combined with an agent (e.g., buffer) that selectively removes (e.g., elutes) either the nucleic acid from the first functional group or the target nucleic acid species from the second functional group of the solid phase carriers, thereby isolating a target nucleic acid species from a mixture comprising a plurality of nucleic acid species.

1 . A kit comprising a solid phase carrier having a surface comprising heterologous first and second functional groups and a cell lysis buffer. 
     
     
         2 . A kit comprising a solid phase carrier having a surface comprising heterologous first and second functional groups and a cell lysis buffer, wherein the solid phase carrier comprises a magnetic microparticle, and wherein the first functional group comprises a COOH functional group. 
     
     
         3 . The kit of  claim 2 , wherein the second functional group comprises an oligo dT functional group, or a derivative thereof. 
     
     
         4 . The kit of  claim 2 , wherein the second functional group comprises an oligonucleotide comprising a sequence that is complementary to a beta globin RNA sequence. 
     
     
         5 . A kit comprising a solid phase carrier having a surface comprising heterologous first and second functional groups, wherein the solid phase carrier comprises a magnetic microparticle, and wherein the first functional group comprises a COOH functional group. 
     
     
         6 . The kit of  claim 5 , wherein the second functional group comprises a streptavidin group. 
     
     
         7 . The kit of  claim 6 , comprising a buffer that is selected from the group consisting of a lysis buffer, a binding buffer, a wash buffer, an elution buffer, or a combination thereof. 
     
     
         8 . The kit of  claim 5 , wherein the second functional group comprises a polymyxin B functional group or a Limulus anti-LPS factor functional group or a derivative thereof. 
     
     
         9 . The kit of  claim 8 , wherein the Limulus anti-LPS factor functional group, or a derivative thereof, is native or recombinant. 
     
     
         10 . The kit of  claim 5 , wherein the second functional group comprises polyethyleneimine or poly-L-lysine or a derivative thereof. 
     
     
         11 . The kit of  claim 5 , wherein the second functional group comprises an oligo dT functional group, or a derivative thereof.