Compositions, methods, and kits for one-step digestion of nucleic acid for analysis by liquid chromatography tandem mass spectrometry
AssigneeUniversity of Cincinnati
InventorsRobert Ross, Patrick A. Limbach, Balasubrahmanyam Addepalli, Manasses Jora
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Abstract
A kit for preparing a nucleic acid sample for analysis by liquid chromatography tandem mass spectrometry (LC-MS/MS) is provided, the kit comprising: a lyophilized enzyme composition comprising: micrococcal nuclease; nuclease P1; and bacterial alkaline phosphatase (BAP); and a digestion buffer. Also provided are enzyme compositions and methods of use for rapid, efficient preparation of a nucleic acid sample for analysis by LC-MS/MS, without the need for denaturation of the sample.
Claims14
A kit for preparing a nucleic acid sample for analysis by liquid chromatography tandem mass spectrometry (LC-MS/MS), the kit comprising:
- a container comprising a lyophilized enzyme composition comprising:
- micrococcal nuclease;
- nuclease P1;
- and
- bacterial alkaline phosphatase (BAP);
- and
- a container comprising a digestion buffer wherein the digestion buffer comprises about 50 mM Tris-HCl at about pH 7.2, about 25 mM Zn2+, and about 5 mM Ca2+.
A method of preparing a nucleic acid sample for analysis by liquid chromatography tandem mass spectrometry (LC-MS/MS), the method comprising:
- (a) providing an enzyme composition comprising:
- micrococcal nuclease;
- nuclease P1;
- bacterial alkaline phosphatase (BAP);
- and a digestion buffer comprising about 50 mM Tris-HCl at about pH 7.2, about 25 mM Zn2+, and about 5 mM Ca2+;
- (b) enzymatically digesting the nucleic acid sample in the enzyme composition to provide a digested nucleoside composition;
- (c) dehydrating the digested nucleoside composition of step (b) to provide a dehydrated digested nucleoside composition;
- and
- (d) rehydrating the dehydrated digested nucleoside composition of step (c) for analysis by LC-MS/MS.
Dependencies are inferred from the claim text.