METHOD FOR THE PURIFICATION OF TARGETED NUCLEIC ACIDS FROM BACKGROUND NUCLEIC ACIDS
The present invention relates generally to the field of nucleic acid purification. In particular, provided herein are micro particles and micro particle clusters for selective anion exchange of nucleic acids, and methods and kits useful for this purpose.
1 . A composition comprising a microparticle having a surface comprising cavities and/or other surface irregularities and/or an aggregate comprising two or more of said microparticles, which aggregate comprises an opening, wherein said surface, cavities, opening, and/or other surface irregularities or pores are: a) functionalized with a weak anion exchange functional group; and b) dimensioned for size exclusion of smaller nucleic acid molecules from larger nucleic acid molecules.
2 . The composition of claim 1 , wherein said larger nucleic molecules are greater than 200 nucleotides in length.
3 . The composition of claim 1 , wherein said larger nucleic acid molecules comprise or are derived from human genomic nucleic acid.
4 . The composition of claim 1 , wherein said smaller nucleic acid molecule comprise or are derived from a microorganism nucleic acid.
5 . The composition of claim 1 , further comprising smaller nucleic acid molecules bound to said pores.
6 . The composition of claim 1 , wherein said microparticle is an iron particle.
7 . The composition of claim 1 , wherein said weak anion exchange functional group is an amine.
8 . The composition of claim 7 , wherein said amino is a primary, secondary, or tertiary alkyl amine.
9 . The composition of claim 7 , wherein said amine has a pKa of greater than 9.
10 . The composition of claim 1 , comprising a plurality of said microparticles.
11 . The composition of claim 10 , wherein said plurality of microparticles are in a resin.
12 . The composition of claim 11 , wherein said resin is on a solid surface.
13 . The composition of claim 12 , wherein said solid surface comprises a plate or column.
14 . The composition of claim 13 , wherein said plate or column comprises a wash buffer.
15 . The composition of claim 14 , wherein said wash buffer is configured to elute said smaller nucleic acid molecules from said microparticle, while leaving behind said larger nucleic acid molecules.
16 . The composition of claim 14 , wherein said wash buffer is compatible with mass spectrometry.
17 . The composition of claim 16 , wherein said wash buffer does not comprise a metal cation salt.
18 . A kit comprising a composition of claim 1 .
19 . The kit of claim 18 , further comprising a wash buffer.
20 . The kit of claim 19 , wherein said wash buffer is configured to elute said smaller nucleic acid molecules from said microparticle, while leaving behind said larger nucleic acid molecules.
21 . The kit of claim 19 , wherein said wash buffer is compatible with mass spectrometry.
22 . The kit of claim 21 , wherein said wash buffer does not comprise a metal cation salt.
23 . A system comprising a composition of claim 1 and an instrument for processing or analyzing a biological sample.
24 . The system of claim 23 , wherein said instrument comprises a nucleic acid amplification device.
25 . The system of claim 23 , wherein said instrument comprises a nucleic acid sequencing device.
26 . The system of claim 23 , wherein said instrument comprises a nucleic acid detection device.
27 . The system of claim 23 , comprising a control computer for automated processing of a plurality of said samples.
28 . A method of detecting a target nucleic acid in a sample comprising: a) exposing a sample to a composition of any of claims 1 ; b) binding smaller nucleic acid from said sample to said pores; c) isolating said smaller nucleic acid by selectively eluting from said pores; and d) detecting said smaller nucleic acid.
29 . The method of claim 28 , wherein said sample is a blood, serum, or plasma sample.
30 . The method of claim 28 , wherein said target nucleic acid is from K. pneunomoniae, E. faecium, S. aureus , or C. albicans.