METHODS FOR REMOVING NUCLEIC ACID CONTAMINATION FROM REAGENTS
In general, the disclosed method can be used to remove contaminating microbes and nucleic acids from microorganisms-derived reagents, apparatus and processes (materials and apparatus) related to PCR (and RT-PCR), including sample prep reagents and materials that are used to isolate, purify and detect nucleic acids.
1 . A method for removing nucleic acid contamination in a PCR master mix reagent comprising:
a. adding a nuclease to the PCR master mix reagent;
b. incubating the PCR master mix reagent plus nuclease; and
c. inactivating the nuclease.
2 . The method of claim 1 , wherein the PCR master mix is formulated for amplifying microorganism nucleic acid.
3 . The method of claim 1 , wherein the nuclease is a DNase or an RNase.
4 . The method of claim 1 , wherein the inactivation of the nuclease is by heat.
5 - 7 . (canceled)
8 . The method of claim 6 , further comprising removal of the nuclease by centrifugation, filtration or magnetic separation.
9 . A method for removing DNA contamination in a solution comprising:
a. passing the solution through a bead, an insoluble matrix, or through a tube, wherein the bead, the insoluble matrix, or the internal surface of said tube comprises at least one DNase; and
b. collecting a DNase treated solution from the bead, the insoluble matrix or the tube.
10 . A method for removing RNA contamination in a solution comprising:
a. adding RNase to the solution;
b. incubating the solution with RNase; and
c. inactivating the RNase.
11 . The method of claim 10 , wherein the RNase is bound to a solid surface.
12 . The method of claim 11 , wherein the solid surface is a bead.
13 . The method of claim 9 , wherein the bead is selected from a group consisting magnetic, non-magnetic, glass and cellulose beads.
14 - 22 . (canceled)
23 . The method of claim 9 wherein said bead or said insoluble matrix is in a column.
24 . The method of claim 9 wherein said DNase is coated or immobilized on the bead or the insoluble matrix.
25 . The method of claim 9 wherein the DNase treated solution is incubated at about 75° C. to produce a resultant solution.
26 . The method of claim 9 wherein the DNase is selected from the group consisting of natural, synthetic or chemically modified DNase.
27 . The method of claim 25 wherein the resultant solution is substantially free of residual DNase activity.
28 . The method of claim 9 wherein the DNase is a recombinant DNase I.
29 . The method of claim 28 wherein the natural DNase is of prokaryotic or eukaryotic origin.
30 . The method of claim 28 wherein the recombinant DNase is TURBO™.
31 . The method of claim 29 wherein the DNase is derived from an organism of either mammalian, nematode, bacterial, yeast, fungal, plant or marine origin.