Reversible association of nucleic acid with a carboxylated substrate

Abstract

The present invention is directed to methods and compositions wherein nucleic acids are associated with a solid phase that comprises a carboxylated substrate. In specific embodiments, precipitation of the nucleic acids occurs in the absence of salt.

1 . A method of isolating nucleic acid, comprising 
 contacting a composition comprising nucleic acid with an effective amount of glycogen to facilitate precipitation of nucleic acid and ethanol in the presence of a carboxylated paramagnetic bead, wherein said contact results in the association of the nucleic acid with the carboxylated paramagnetic bead.    
   
   
       2 . The method of  claim 1 , wherein said nucleic acid is subsequently eluted from the paramagnetic bead.  
   
   
       3 . The method of  claim 1 , wherein precipitation occurs in the substantial absence of salt.  
   
   
       4 . The method of  claim 1  wherein an effective amount of glycogen comprises from about 1 μg/mL to about 1000 μg/mL.  
   
   
       5 . The method of  claim 4 , wherein an effective amount of glycogen comprises from about 200 μg/mL to about 500 μg/mL.  
   
   
       6 . The method of  claim 5 , wherein an effective amount of glycogen comprises about 250 μg/mL.  
   
   
       7 . The method of  claim 1 , wherein said nucleic acid is DNA.  
   
   
       8 . The method of  claim 7 , wherein the DNA is genomic DNA.  
   
   
       9 . The method of  claim 7 , wherein the DNA is plasmid DNA.  
   
   
       10 . The method of  claim 7 , wherein said DNA is an oligonucleotide.  
   
   
       11 . The method of  claim 1 , wherein said nucleic acid is RNA.  
   
   
       12 . The method of  claim 1 , wherein said nucleic acid is from 1-100 nucleotides in length.  
   
   
       13 . The method of  claim 1 , wherein said nucleic acid is from 100-1000 nucleotides in length.  
   
   
       14 . The method of  claim 1 , wherein said nucleic acid is from 1000-10,000 nucleotides in length.  
   
   
       15 . The method of  claim 1 , wherein said nucleic acid is from 10,000-1,000,000 nucleotides in length.  
   
   
       16 . A method for isolating nucleic acid, comprising 
 contacting a composition comprising nucleic acid with an effective amount of glycogen to facilitate precipitation of nucleic acid and ethanol in the presence of a carboxylated substrate, wherein said contact results in the association of the nucleic acid with the carboxylated substrate.    
   
   
       17 . The method of  claim 16 , wherein said nucleic acid is subsequently eluted from the carboxylated substrate.  
   
   
       18 . The method of  claim 16 , wherein said method is carried out in the substantial absence of salt.  
   
   
       19 . The method of  claim 16 , wherein said nucleic acid is DNA.  
   
   
       20 . The method of  claim 19 , wherein the DNA is genomic DNA.  
   
   
       21 . The method of  claim 19 , wherein the DNA is plasmid DNA.  
   
   
       22 . The method of  claim 19 , wherein said DNA is an oligonucleotide.  
   
   
       23 . The method of  claim 16 , wherein said nucleic acid is RNA.  
   
   
       24 . The method of  claim 16 , wherein said nucleic acid is from 1-100 nucleotides in length.  
   
   
       25 . The method of  claim 16 , wherein said nucleic acid is from 100-1000 nucleotides in length.  
   
   
       26 . The method of  claim 16 , wherein said nucleic acid is from 1000-10,000 nucleotides in length.  
   
   
       27 . The method of  claim 16 , wherein said nucleic acid is from 10,000-1,000,000 nucleotides in length.  
   
   
       28 . A kit for nucleic acid isolation, comprising in a suitable containing means: 
 a carboxylated substrate; and    a binding solution containing glycogen.    
   
   
       29 . A composition for reversibly binding nucleic acid to a carboxylated substrate, comprising: 
 ethanol;    a nucleic acid;    an effective amount of glycogen to facilitate precipitation of nucleic acid; and    a carboxylated substrate.    
   
   
       30 . The composition of  claim 29 , wherein said composition substantially lacks salt.  
   
   
       31 . The composition of  claim 29 , wherein said carboxylated substrate is a paramagnetic bead.  
   
   
       32 . The composition of  claim 29 , wherein said nucleic acid is DNA.  
   
   
       33 . The composition of  claim 32 , wherein the DNA is genomic DNA.  
   
   
       34 . The composition of  claim 32 , wherein the DNA is plasmid DNA.  
   
   
       35 . The composition of  claim 32 , wherein said DNA is an oligonucleotide.  
   
   
       36 . The composition of  claim 29 , wherein said nucleic acid is RNA.  
   
   
       37 . The composition of  claim 29 , wherein said nucleic acid is from 1-100 nucleotides in length.  
   
   
       38 . The composition of  claim 29 , wherein said nucleic acid is from 100-1000 nucleotides in length.  
   
   
       39 . The composition of  claim 29 , wherein said nucleic acid is from 1000-10,000 nucleotides in length.  
   
   
       40 . The composition of  claim 29 , wherein said nucleic acid is from 10,000-1,000,000 nucleotides in length.  
   
   
       41 . An instrument for isolating nucleic acid, comprising: 
 a means for contacting a composition comprising nucleic acid with an effective amount of glycogen to facilitate precipitation of nucleic acid and ethanol in the presence of a carboxylated substrate, wherein said contact results in the association of the nucleic acid with the carboxylated substrate.    
   
   
       42 . An instrument for isolating nucleic acid comprising: 
 means for containing a nucleic acid sample;    means for adding ethanol to the nucleic acid sample    means for adding glycogen to the nucleic acid sample in an amount effective to cause precipitation of the nucleic acid;    a carboxylated substrate in contact with the nucleic acid sample.    
   
   
       43 . The instrument of  claim 42 , wherein the means for containing the nucleic acid sample is a plate containing individual wells.  
   
   
       44 . The instrument of  claim 42 , wherein the means for containing the nucleic acid sample is a holder for a plate containing individual wells.  
   
   
       45 . The instrument of  claim 42 , wherein the means for containing the nucleic acid sample is a holder for centrifuge tubes.  
   
   
       46 . The instrument of  claim 42 , further comprising means for removing the carboxylated substrate from the sample.  
   
   
       47 . The instrument of  claim 46 , wherein the carboxylated substrate is paramagnetic beads and the means for removing the substrate from the sample comprises a means for generating an electric field.  
   
   
       48 . The instrument of  claim 42 , wherein the carboxylated substrate in contact with the nucleic acid sample is the inner surface of a centrifuge tube.